LECTURE ON BIOLOGICAL CHEMISTRY FOR 2 YEAR STUDENTS

OF MEDICAL FACULTY ( 2005- 2006)

LECTURE 24
THEME: BIOCHEMISTRY OF BLOOD, MUSCLE AND CONNECTIVE
TISSUE
!LAN OF LECTURE
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"" Mechanism of contraction and relaxation. Role of Ca
2+
.
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The blood is the most specialized fluid tissue which circulates in vascular
system and together with lymph and intercellular space compounds an internal
environment of an organism.
The blood executes such functions:
1. Transport of gases oxygen from lungs is carried to tissues and carbon
dioxide from tissues to lungs.
2. Transport of nutrients to all cells of organism (glucose, amino acids, fatty
acids, vitamins, etone bodies, trace substances and others!. "ubstances such as urea,
uric acid, bilirubin and creatinine are taen away from the different organs for
ultimate excretion.
#. $egulatory or hormonal function hormones are secreted in to blood and
they are transported by blood to their target cells.
%. Thermoregulation function & an exchange of heat between tissues and blood.
'. (smotic function& sustains osmotic pressure in vessels.
). *rotective function& by the phagocytic action of leucocytes and by the actions
of antibodies, the blood provides the most important defense mechanism.
+. ,etoxification function & neutralization of toxic substances which is
connected with their decomposition by the help of blood enzymes.
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Two types of blood cells can be distinguished & white and red blood cells.
-hite blood cells are called leucocytes. Their .uantity in adult is %&/ x 10
/
12.
$ed blood cells are called erythrocytes. Their .uantity in peripheral blood is %,'&
' x 10
12
12. 3esides that, there are also thrombocytes or platelets in blood.
2eucocytes (white blood cells! protect an organism from microorganisms,
viruses and foreign substances, that provides the immune status of an organism.
2eucocytes are divided into two groups4 5ranulocytes and agranulocytes.
5ranulocytes consist of neutrophils, eosinophils and basophils. 6granulocytes consist
of monocytes and lymphocytes.
Neutrophils
7eutrophils comprise of )0&+0 8 from all leucocytes. Their main function is to
protect organisms from microorganisms and viruses. 7eutrophils have segmented
nucleus, endoplasmic reticulum (underdeveloped! which does not contain ribosomes,
insufficient amount of mitochondria, well&developed 5olgi apparatus and hundreds of
different vesicles which contain peroxidases and hydrolases. (ptimum condition for
their activity is acidic p9. There are also small vesicles which contain alaline
phosphatases, lysozymes, lactopherins and proteins of cationic origin.
5lucose is the main source of energy for neutrophils. :t is directly utilized or
converted into glycogen. /0 8 of energy is formed in glycolysis, a small amount of
glucose is converted in pentosophosphate pathway. 6ctivation of proteolysis during
phagocytosis as well as reduction of phosphatidic acid and phosphoglycerols are also
observed. The englobement is accompanied by intensifying of a glycolysis and
pentosophosphate pathway. 3ut especially intensity of absorption of oxygen for
neutrophils & so&called flashout of respiration grows. 6bsorbed oxygen is spent for
formation of its fissile forms that is carried out with participation enzymes4
1. 76,*;< &(=>,6"? catalyzes formation of super oxide anion in reaction

2. 6n enzyme 76,9& (=>,6"? is responsible for formation of hydrogen
peroxide
#. @yeloperoxydase catalyzes formation of hypochloric acid from chloride and
hydrogen peroxide
6nion of hypochloric acid may react with the following molecule of hydrogen
peroxide and form singlet oxygen.
The active forms of oxygen show bactericidal activity and destroy microbial
nucleic acids, proteins and lipids. The leading role in bactericidal activity of
leucocytes belongs to hydrogen peroxide and hypochloride. Aormed hypochloride
produces a chlorination of frames of a microbial membrane that is accompanied by
their destruction. Byeloperoxydase with the help of hypochloride decarboxylates
amino acids.
:n the function of phagocytosis neutrophils are also assisted by leuotrienes
(arachidonic acid derivatives! by means of chemotaxis stimulation.
Thus, in the phagocytosis of neutrophils participate many elements of
fermentative and non&fermentative character and with different activity mechanism.
Basophiles
3asophiles mae up 1&'8 of all blood leuocytes. They are actively formed in
the bone marrow during allergy. 3asophiles tae part in the allergic reactions, in the
blood coagulation and intravascular lipolysis. They have the protein synthesis
mechanism, which wors due to the biological oxidation energy . They synthesize the
mediators of allergic reactions histamine and serotonin, which during allergy cause
local inflammation. 9eparin, which is formed in the basophiles, prevents the blood
coagulation and activates intravascular lipoprotein lipase, which splits
triacylglycerin.
Eosinophiles
They mae up #&)8 of all leuocytes. ?osinophiles as well as neutrophiles
defend the cells from microorganisms, they contain myeloperoxidase, lysosomal
hydrolases. 6bout the relations of eosinophiles with testifies the growth of their
amount during the sensitization of organism, i.e. during bronchial asthma,
helminthiasis. They are able to pile and splits histamine, Cto dissolveD thrombus with
the participation of plasminogen and bradyinin&ininase.
Monocytes
They are formed in the bone marrow. They mae up %&E8 of all leuocytes.
6ccording to the function they are called macrophages. Tissue macrophages derive
from blood monocytes. ,epending on their position they are called4 in the liver
reticuloendotheliocytes, in the lungs & alveolar macrophages, in the intermediate
substance of connective tissue histocytes etc. Bonocytes are characterized by a
wide set of lysosomal enzymes with the optimum activity in the acidic condition.
The maFor functions of monocytes and macrophages are endocytosis and
phagocytosis.
Lymphocytes
The amount 20&2'8, are formed in the lymphoid tissue or thymus, play important
role in the formation of humoral and cellular immunity. 2ymphocytes have powerful
system of synthesis of antibody proteins, energy is maForily pertained due to
glycolysis, rarely by aerobic way.
Thrombocytes (blood platelets)
The amount less than 18, they play the main role in the process of hemostasis.
They are formed as a result of disintegration of megaaryocytes in the bone marrow.
Their life&time is +&/ days. :n spite of the fact that thrombocytes have no nucleus,
they are able to perform practically all functions of the cell, besides ,76 synthesis.
Erythrocytes
9uman blood contains 2' trillion of erythrocytes. Their main function
transportation of (
2
and G(
2
they perform due to the fact that they contain #%8 of
hemoglobin, and per dry cells mass /'8. The total amount of hemoglobin in the
blood e.uals 1#0&1)0 g1l. :n the process of erythropoesis the preceding cells decrease
their size. Their nuclei at the end of the process are ruined and pushed out of the cells.
/08 of glucose in the erythrocytes is decomposed in the process of glycolysis and
108 & by pentose&phosphate way. There are noted congenital defects of enzymes of
these metabolic ways of erythrocytes. ,uring this are usually observed hemolytic
anemia and other structural and functional erythrocytesH affections.
BLOOD !LASMA !ROTEINS
6. *rotein fractions which are received by the electrophoresis
3.
*rotein fractions which are received with the help of imunoelectropheresis on agar
gel.
*rotein Goncentration
6cidic I
1
glycoproteid 0,20 0,%0 g1l
I
1
6ntitrypsyn 2,00&%,00 g1l
Geruloplasmin 0,1'&0,)0 g1l
Gu
2J
1),0&#1,0 mmmol1l
9aptoglobine 1,00&%,00 g1l
IK
2
& Bacroglobulin 2,'0&#,'0 g1l
Transpheryn 2,'0&%,10 g1l
Ae
#J
11,0&2+,0 mmmol1l
Aibrinogen 2,00&%,00 g1l
Aractions Goncentration $elative contents
6lbumin #E,0 & '0,0 g1l 0,'0 & 0,)0
I
1
globulins 1,% #,0 g1l 0,01 & 0,0'
IK
2
globulins ',) /,1 g1l 0,0+ & 0,1#
L& globulins ',% /,1 g1l 0,0/ 0,1'
M globulins /,1 1%,+ g1l 0,1% 0,22
Total protein )',0 E', 0 g1l 1,00
:mmunoglobulins (:g!
:g5 E,00&1E,00 g1l
:g6 1,00&%,00 g1l
:gB 0,)0&2,E0 g1l
:g, 0,00&0,1' g1l
:g? Till 'x10
&%
6lbumin is the maFor constituent (''&)08! of plasma proteins. 9uman albumin
has comparatively low molecular weight of ''000 & +0000 , and less molecular size
(1#;# nm!, than globulins and fibrinogen. *lasma albumin is synthesized in the liver
(10&1' g per day!. :ts main functions are4
1! osmotic blood pressure maintenance, thus participation in the regulation of water
distribution between blood and inter&cellular spaceN
2! transport functionN
#! detoxification function.
?lectrophoresis separates globulins into
1
&,
2
&, & and &globulins. ?ach of the
fractions includes great amount of individual proteins. +'&/08 of &globulins and
'08 of &globulins are synthesized by hepatocytes. & and &globulins are transport
proteins4 vitamin 6 is transported by retinol&binding protein, thyroxine thyroxine&
binding protein, transcortin is the transporter of hormones cortisol and
corticosterone, ceruloplasmin copper ions, transferin iron ions. The inhibitors of
proteolytic enzymes are alpha&1&antitrypsin, alpha&2&macroglobulin, inter&a&trypsin
inhibitor. 5lobulins haptoglobin and hemopexin prevent loss of &heme iron with
urine. 9aptoglobins (alpha&2&globulin fraction! bond with hemoglobin attenuated in
the plasma and thier complexes are pertained by reticulendothelium cells, where are
decomposed. 6nalogically hemopexin (&globulin fraction! and heme complex are
pertained by the livers. Aree iron is reutilized. & and &globulin fractions include
lipoproteins, observed in the part C2ipidsD. 9,2* or &2* migrate in the
electrophoresis togrther with &globulinsN 2,2* or &2* with &globulinsN O2,2*
or pre&&2* between & and &lipoproteinsN chylomicrons not move in the electric
field and stay in the start position.
&globulin fraction contains, maForily, antibodies (immunoglobulins!. 6ntibodies
are synthesized by 3&lymphocytes. The amount of individual antibodies, different by
primary structure, is extremely big. Thus in the organism of a single man can be
synthesized up to 10
+
different antibodies. The structure, functions and synthesis of
antibodies are observed in the part C:mmunochemistryD.
To the fraction of antibodies belong also pathological proteins, synthesized by
myeloma by specific cells of antibodies&forming system and appear in great amount
in the plasma of the patients. "uch myeloma globulins are fragments of antibodies
and can be filtered in the idneys and secreted with urine. They are also called 3ence
Pones protein. :ts peculiarity is sediment formation in the acidic medium at '0&)0
0
G
and redissolution at higher temperature.
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Gells are the basis units of life. Bost mammalian cells are located in tissues,
where they are surrounded by a complex extracellular matrix (ECM!, often referred
to as connective tissue. This matrix has a variety of important functions, apart from
acting as a supporting scaffolding for the cells it surrounds.
?GB contains # maFor classes of biomolecules4
1. The structural proteins, collagen and elastinN
2. Gertain specialized proteins, such as fibrillin, fibronectin, and lamilin,
which have specific functions in the ?GBN
#. *roteoglycans, which consist of long chains of repeating disaccharides
(glycosaminoglycans, 565s, formerly called mucopolysaccharides! atached to
specific core proteins.
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Gollagen, the maFor component of most connective tissues, constitutes
approximately 2' 8 of the protein of mammals. :t provides an extracellular
framewor for all metazoan animals and exists in virtually every animal tissue. Bore
than 12 distinct types of collagen have been identified in mammalian tissues.
?xamples of collagens types4
type 2ocalization "et of polypeptide chains
: "in, bones, tendon, cornea of eye,
sclera
[
1(:!
]
2

2
:: Gartilages, vitreous body
[
1
(::!]
#
:::
"in of fetus, walls of large blood
vessels
[
1
(:::!]
#
:O basal membrane
[
1
(:O!]
#
6lthough several of these are present only in small proportions, thay may play
important roles in determining the physical properties of the tissues.
6ll collagen types have a triple helical structure. ?ach polypeptide subunit or
alpha chain is twisted into a left&handed helix of # residues per turn. Three of these
alpha chains are then wound into a right&handed super&helix, forming a rodlie
molecule 1.% nm in diameter and about #00 nm long. 6 striing characteristic of
collagen is the the occurrence of glycine residues at every third position of the triple
helical portion of the alpha chain. This is necessary because glycine is the only amino
acid small enough to be accomodated in the limited space available down the central
core of the triple helix. This repeating structure, represented as (G%4->-Y!n, is an
absolute re.uirement for the formation of the triple helix. -hile > and Y can be any
other amino acids, about 100 of the > position are proline and about 100 Y position
are hydroxyproline. 9ydroxyproline is formed by the posttranslational hydroxylation
of peptide&bound proline residues catalyzed by the enzyme prolyl hydroxylase, whose
cofactors are ascorbic acid and I&etoglutarate. 2ysines in the Y position may also be
posttranslationally modified to hydroxylysine through the action of lysyl
hydroxylase, an enzyme with similar cofactors. "ome of these hydroxylysines may be
further modified by the addition of galactose or galactosyl&glucose through an (&
glycosidic linage, a glycosylation site that is uni.ue to collagen.
Gollagen types that form long rodlie fibers in tissues are assembled by lateral
association of these triple helical units into a C.uarter&staggeredD alignment such that
each is displaced longitudinally from its neighbor by slightly less than one&.uarter of
its length. This arrangement is responsible for the banded appearance of these fibers
in connective tissues. Gollagen fibers are further stabilized by the formation of
covalent cross&lins, both within and between the triple helical units. These cross&
lins from through the action of lysyl oxidase, a copper&dependent enzyme that
oxidatively deaminates the Q&amino groups of certain lysine and hydroxylysine
residues, yielding reactive aldehydes. "uch aldehydes can form aldol condensation
products with other lysine& or hydroxylysine&derived aldehydes, or from "chiff bases
with the Q&amino groups of unoxidized lusines or hydroxylysines. These reactions,
after further chemical rearrangements, result in the stable, covalent cross&lins that
are important for the tensile strength of the fibers.
"everal collagen types do not form banded fibers in tissues. 6t the molecular
level, these collagens are characterized by interruptions of the triple helix with
stretches of protein lacing G%4->-Y repeat se.uences. These +$+-G%4->-Y
se.uences result in areas of globular structure interspersed in the triple helical
structure. T46/ IV collagen, the best characterized example of collagens with
discontinuous triple helices, is an important component of basement membranes
where it forms a meshlie networ.
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7ewly synthesized collagen undergoes extensive posttranslational modification
before becoming part of a mature, extracellular collagen fiber. 2ie most secreted
proteins, collagen is synthesized on ribosomes in a precursor form, preprocollagen,
which contains a leader or signal se.uence that directs the polypeptide chain into the
vesicular space of the endoplasmatic reticulum. 6s it enters the endoplasmatic
reticulum, this leader se.uence is enzymatically removed. 9ydroxylation of proline
and lysine residues and glycosylation of hydroxylysines in this procollagen molecule
also tae place at this time. The procollagen molecule contains polypeptide
extensions of 20,000&#',000 B- at both its amino& and carboxy&terminal ends,
neither of which is present in mature collagen. 3oth extension peptides contain
cysteine residues. -hile the aminoterminal propeptide forms only intrachain disulfide
bonds, the carboxy&terminal propeptides form both intrachain and interchain disulfide
bonds. Aormation of these disulfide bonds assists in the registration of the # collagen
molecules to form the triple helix, winding from the carboxyl&terminal end. 6fter
formation of the triple helix, no further hydroxylation of proline or lysine or
glycosylation of hydroxylysines can tae place.
Aollowing secretion from the cell by way of the 5olgi apparatus, extracellular
enzymes called procollagen aminoproteinase and procollagen carboxyproteinase
remove the extension peptides at the amino& and carboxy&terminal ends, respectively.
Gleavage of these propetides may occur within crypts or folds in the cell membrane.
(nce the propeptides are removed, the triple helical collagen molecules, containing
approximately 1000 amino acids per chain, spontaneously assemble into collagen
fibers. These are further stabilized by the formation of inter& and intrachain cross&
lins through the action of lysyl oxidase, as described previously.
The same cells that secrete collagen also secrete fibronectin, a large
glycoprotein present on cell surfaces, in the extracellular matrix, and in blood.
Aibronectin binds to aggregating precollagen fibers and alters the inetics of fiber
formation in the pericellular matrix. 6ssociated with fibronectin and procollagen in
this matrix are the proteoglycans heparan sulfate and chondroitin sulfate. :n fact, type
:= collagen, a minor collagen type from cartilage, contains attached proteoglycan
chains. "uch interactions may serve to regulate the formation of collagen fibers and
to determine their orientation in tissues.
E%(-,#+ 9 2(#+ 63$,/#+ $) /%(-,#' )#03#%-, -,3*',*3/ (+1 0#$%$&#'(% 3$%/.
?lastin is a connective tissue protein that is responsible for properties of
extensibility and elastic recoil in tissues. 6lthough not as widespread as collagen,
elastin is present in large amounts, particularly in tissues that re.uire these physical
properties, eg, lung, large arterial blood vessels, and some elastic ligaments. "maller
.uantities of elastin are also found in sin, ear cartilage, and several other tissues. :n
contrast to collagen, there appears to be only one genetic type of elastin, although
variants arise by differential processing of the hn$76 for elastin. ?lastin is
synthesized as a soluble monomer of +0,000 B- called tropoelastin. "ome of the
prolines of tropoelastin are hydroxylated to hydroxyproline by prolyl hydroxylase,
although hydroxylysine and glycosylated hydroxylysine are not present. Rnlie
collagen, tropoelastin is not synthesized in a pro& form with extension peptides.
Aurthermore, elastin does not contain repeat 5ly&=&> se.uences, triple helical
structure, or carbohydrate moieties.
6fter secretion from the cell, certain lysyl residues of tropoelastin are
oxidatively deaminated to aldehydes by lysyl oxidase, the same enzyme involved in
this process in collagen. 9owever, the maFor cross&lins formed in elastin are the
desmosines, which result from the condensation of # of these lysine&derived
aldehydes with an unmodified lysine to form a tetrafunctional cross&lin uni.ue to
elastin. (nce cross&lined in its mature, extracellular form, elastin is highly insoluble
and extremely stable and has a very low turnover rate. ?lastin exhibits a variety of
random coil conformations that permit the protein to stretch and subse.uently recoil
during the performance of its physiologic functions.
S,3*',*3/ (+1 )*+',#$+- $) 63$,/$&%4'(+-.
*roteoglycans are proteins that contain covalently lined glycosaminoglycans
(565s!. The protein bound covalently to 565s are called core proteinsN they have
proved difficult to isolate and characterize. The amount of carbohydrate in a
proteoglycan is usually much greater than is found in a glycoprotein and may
comprise up to /' 8 of its weight. There are at least + 565s4 hyaluronic acid,
chondroitin sulfate : and ::, heparin, heparan sulfate, and dermatan sulfate. 6 565 is
an unbranched polysaccharide made up of repeating disaccharides, one component of
which is always an amino sugar (hence the name 565!, either ,&glucosamine or ,&
galactosamine. The order component of the repeating disaccharides (except in the
case of eratan sulfate! is a uronic acid, either 2&iduronic acid (:dR6!. -ith the
exception of hyaluronic acid, all the 565s contain sulfate groups, either as (&esters
or as 7&sulfate (in heparin and heparan sulfate!. 9yaluronic acid affords another
exception because there is no clear evidence that it is attached covalently to protein,
as the definition of a proteoglycan given above specifies.
*roteoglycans are found in every tissue of the body, mainly in the extracellular
matrix. There they are associated with each other and some of them with collagen,
another with elastin. These interactions are important in determining the structural
organization of the matrix. :n addition, some of them interact with certain adhesive
proteins, such as fibronectin and laminin. The 565s present in the proteoglycans are
polyanions and hence bind polycations and cations such as 7a and S. This latter
ability attracts water by osmotic pressure into the extracellular matrix and contributes
to its turgor. 565s also gel at relatively low concentrations. 3ecause of the long
extended nature of the polysaccharide chain of 565s and their ability to gel, the
proteoglycans can act as sieves, restricting the passage of large macromolecules into
the extracellular matrix but allowing relatively free diffusion of small molecules.
6gain, because of their extended structures and the huge macromolecular aggregates
that they often form, they occupy a very large volume of the matrix relative to
proteins.
The + 565s named above differ from each other in a number of the following
properties4 amino sugar composition, uronic acid composition, linages between
these components, chain length of the disaccharides, presence or absence of sulfate
groups and their positions of attachment to the constituent sugars, nature of the core
proteins to which they are attached, nature of the linage to core protein, their tissue
and subcellular distribution, and their biologic functions.
H4(%*3$+#' A'#1: consists of an unbranched chain of repeating disaccharide
units containing 5lcR6 and 5lc76c. There is no firm evidence that it is lined to
protein, as are the other 565s. 9yaluronic acid is present in bacteria and is widely
distributed among various animals and tissues, including synovial fluid, the vitreous
body of the eye, and loose connective tissue. 6lso it present in high concentration in
embryonic tissues and is thought to play an important role in permitting cell
migration during morphogenesis and wound repair. :ts ability to attract water into the
extracellular matrix and thereby Cloosen it upD may be important in this regard. The
high concentrations of hyaluronic acid and chondroitin sulfates present in cartilage
contribute to its compressibility.
C.$+13$#,#+ S*%)(,/- (Ghondroitin %&"ulfate and Ghondroitin )&"ulfate!4
proteoglycans lined to chondroitin sulfate by the =yl&"er (&glycosidic bond are
prominent components of cartilage. The repeating disaccharide is similar to that
found in hyaluronic acid, containing 5lcR6 but with 5al76c replacing 5lc76c. The
5al76c is substituted with sulfate at either its % or its ) position, with approximately
one sulfate being present per disaccharide unit. ?ach chain contains some %0
disaccharide units and thus has a B- of about 20,000. Bany such chains are
attached to a single protein molecule, generating massive proteoglycans of high B-
(eg, in nasal cartilage approximately 2.'T10
)
!. The chondroitin sulfate associate
tightly with hyaluronic acid with the aid of 2 Clin proteinsD that bind
hydrophobically to both hyaluronic acid and to the core protein, generating very large
aggregates in connective tissue. The chondroitin sulfates are located at sites of
calcification in endochondral bone. 6lso this proteoglycan is located inside certain
neurons and may provide an endoseletal structure, helping to maintain their shape.
?/3(,(+ S*%)(,/- I (+1 II4 this substances consist of repeating 5al&5lc&7ac
disaccharide units containing sulfate attached to the ) position of 5lc76c or
occasionally of 5al. Type : is abundant in cornea, and type :: is found along with
chondroitin sulfate attached to hyaluronic acid in loose connective tissue. Types : and
:: have different attachments to protein.
H/6(3#+: the repeating disaccharide contains glucosamine (5lc7! and either of
the 2 uronic acids. Bost of the amino groups of the 5lc7 resirues are 7&sulfated, but
a few are acetylated. The 5lc7 also carries a G
)
sulfate ester. 6pproximately /0 8 of
the uronic acid residues are :dR6. :nitially, all of the uronic acids are 5lcR6, but a '&
epimerase converts approximately /0 8 of the 5lcR6 residues to :dR6 after the
polysaccharide chain is formed. The protein molecule of the heparin proteoglycan is
uni.ue, consisting exclusively of serine and glycine residues. 6pproximately two&
thirds of the serine residues contain 565 chains, usually with an B- of '000&1'000
but occasionally much higher. 9eparin is found in the granules of mast cells and also
in liver, lung, and sin. 9eparin is an important anticoagulant. :t binds with factors :=
and =:, and also interact with plasma antitrombin :::.
H/6(3(+ S*%)(,/4 this molecule is present on many cell surfaces as a
proteoglycan and is extracellular. :t contains 5lc7 with fewer 7&sulfates than
heparin, and unlie heparin, its predominant uronic acid is 5lcR6. :t plays important
role in the mediation of cell growth and cell&cell communication.
D/32(,(+ S*%)(,/4 this is widely distributed in animal tissues. "tructurally, it
resembles both the chondroitin sulfate and heparan sulfate. :ts structure is similar to
that of chondroitin sulfate, except that in place of a 5lcR6 in L&1,# linage to
5al76c, it contains an :dR6 in an I&1,# linage to 5al76c. Aormation of the :dR6
occurs, as in heparin and heparan sulfate, by '&epimerization of 5lcR6. 3ecause this
is regulated by the degree of sulfation, and sulfation is incomplete, dermatan sulfate
contains both :dR6&5al76c and 5lcR6&5al76c disaccharides.
S,3*',*3(% (+1 './2#'(% 6/'*%#(3#,#/- $) ,./ 2*-'%/.
Buscle is the maFor biochemical transducer (machine! that converts potential
(chemical! energy into inetic (mechanical! energy. Buscle, the largest single tissue
in the human body, maes up somewhat less than 2' 8 of body mass at birth, more
than %0 8 in the young adult, and somewhat less than #0 8 in the ages adult.
6n effective chemical&mechanical transducer must meet several re.uirements4
1. There must exist a constant supply of chemical energy. :n vertebrate muscle,
6T* and creatine phosphate supply chemical energy.
2. There must be a means of regulating the mechanical activity ie, the speed,
duration, and force of contraction in the case of muscle.
#. The machine must be connected to an operator, a re.uirement met in
biologic systems by the nervous system.
%. There must be a way of returning the machine to its original state.
Buscle is a pulling, not a pushing, machine. Therefore, a given muscle must be
antagonized by another group of muscles or another force such as gravity or elastic
recoil.
:n vertebrates, the above re.uirements and the specific needs of the organisms
are met by # types of muscles4 seletal muscle, cardiac muscle, and smooth muscle.
3oth seletal and cardiac muscle appear striated upon microscopic observationN
smooth muscle is nonstriated. 6lthough seletal muscle is under voluntary nervous
control, the control of both cardiac and smooth muscle is involuntary.
"triated muscle is composed of multinucleated muscle fiber cells surrounded
by an electrically excitable membrane, the sarcolemma. 6n individual muscle fiber
cell, which may extend the entire length of the muscle, contains a bundle of many
myofibrils arranged in parallel, embedded in intracellular fluid termed sarcoplasm.
-ithin this fluid is contained glycogen, the high&energy compounds 6T* and
phosphocreatine, and the enzymes of glycolysis.
The sarcomere is repeated along the axis of a fibril at distance of 1'00&2#00
nm. -hen the myofibril is examined by electron microscope, alternating dar and
light bands (6 bands and : bands! can be observed. The central region of the 6 band
(the 9 zone! appears less dense than the rest of the band. The : band is bisected by a
very dense and narrow U line.
-hen myofibrils are examined by electron microscopy, it appears that each
myofibril is constructed of 2 types of longitudinal filaments. (ne type, the thic
filament, confined to the 6 band, contains chiefly the protein myosin. These filaments
are about 1) nm in diameter and arranged in cross section as a hexagonal array.
The other filament, the thin filament, lies in the : band and extends also into
the 6 band but not into the 9 zone of the 6 band. The thin filaments contain the
proteins actin, tropomyosin, and troponin. :n the 6 band, the thin filaments are
arranged around the thic (myosin! filaments as a secondary hexagonal array. ?ach
thin filament lies symmetrically between # thic filaments, and each thic filament is
surrounded symmetrically by ) thin filaments.
!3$,/#+- $) -(3'$6%(-2(
Byogen group of the some glycolysis enzymes.
Byoglobin chromoprotein lie hemoglobin. Gontains Ae
2J
, bonds to oxygen
about ' times more strongly than does hemoglobin and provides (
2
for oxidative
processes and aerobic glycolysis.
Byoalbumin group of lie blood albumin proteins.
!3$,/#+- $) 24$)#03#%-
Bonomeric (globular! actin (5&actin! is a %#,000&B- globular protein that
maes up 2' 8 of muscle protein by weight. 6t physiologic ionic strength and in the
presence of magnesium, 5&actin polymerizes noncovalently to form an insoluble
double helical filament called A&actin.The A&actin fiber is )&+ nm thic and has a
pitch or repeating structure every #'.' nm. 7either 5& nor A&actin exhibits any
catalytic activity.
Byosin contributes '' 8 of muscle protein by weight and forms the thic
filaments. Byosin is an asymmetric hexamer with a molecular weight of %)0,000.
The myosin has a fibrous portion consisting of 2 interwined helices, each with a
globular head portion attached at one end. The hexamer consists of one of heavy
chains and 2 pair of light chains. "eletal muscle myosin exhibits 6T*&hydrolyzing
(6T*&ase! activity and binds to A&actin, an insoluble molecule.
:n striated muscle, there are 2 other proteins that are minor in terms of their
mass but important in terms of their function. Tropomyosin is a fibrous molecule that
consists of 2 chains, alpha and beta, that attach to A&actin in the groove between its
filaments. Tropomyosin is present in all muscular and muscle&lie structures.
The troponin complex is uni.ue to striated muscle and consists of #
polypeptides. Troponin T (TpT! binds to tropomyosin as well as to the other 2
troponin components. Troponin : (Tp:! inhibits the A&actin&myosin interaction and
also binds to the other components of troponin. Troponin G (TpG! is a calcium&
binding polypeptide that is structurally and functionally analogous to calmodulin, an
important calcium&binding protein widely distributed in nature. Aour molecules of
calcium ion are bound per molecule of troponin G pr calmodulin, and both molecules
have a molecular weight of 1+,000.
Stromal proteins
:n seletal muscles stromal proteins presented by collagen, neuroeratin,
elastin etc.
Mechanism of contraction and relaxation. Role of Ca
2+
.
-hen muscle contracts, there is no change in the lengths of the thic filaments
or of the thin filaments, but 9 zone and the : bands shorten. Thus, the arrays of
interdigitating filaments must slide past one another during muscle contraction. The
crossbridges generate and sustain the tension.
Buscle contraction consists of the cyclic attachment and detachment of the
globular head portion of myosin to the A&actin. The attachment is followed by a
change in the actin&myosin interaction, so that the actin filaments and the myosin
filaments slide past one another. The energy is supplied indirectly by 6T*.
The biochemical cycle of muscle contraction consists of ' steps4 (1! The
myosin head alone can hydrolyze 6T* to 6,* J *
i
, but it cannot release the products
of this hydrolysis. Thus, the hydrolysis of 6T* by the myosin head alone is
stoichiometric rather than catalytic. (2! The myosin head containing 6,* and *
i
can
rotate freely through large angles in order to locate and bind to A&actin, maing an
angle of about /0 degrees with the fiber axis. This interaction (#! promotes the
release of 6,* and *
i
from the actin&myosin complex. 3ecause the conformation of
lowest energy for the actomyosin bond is %' degrees, the myosin changes its angle
from /0 degrees to about %' degrees by pulling the actin (10&1' nm! toward the
center of the sarcomere. (%! 6 new 6T* molecule binds to the myosin&A&actin
complex. Byosin&6T* has a poor affinity for actin, and thus the myosin (6T*! head
is released ('! from the A&actin. This last step is relaxation, a process clearly
dependent upon the binding of 6T* to the actin&myosin complex. The 6T* is again
hydrolyzed by the myosin head but without releasing 6,* J *
i
, to continue the cycle.
:n resting muscle sarcoplasm, the concentration of Ga
2J
is 10&+&10&E mol12.
The resting state is achieved because Ga
2J
is pumped into the sarcoplasmic reticulum
through the action of an active transport system, called Ga
2J
6T*ase, initiating
relaxation. The sarcoplasmic reticulum is a networ of fine membranous sacs. :nside
the sarcoplasmic reticulum, Ga
2J
is bound to a specific Ga
2J
&binding protein
designated calse.uestrin. The sarcomere is surrounded by an excitable membrane (the
T tubule sustem! composed of transverse (T! channels closely associated with the
sarcoplasmic reticulum. -hen the sarcolemma (ie, the plasma membrane of the
muscle cell! is excited by a nerve impulse, the signal is transmitted into the T tubule
system and a Ga
2J
release channel in the nearby sarcoplasmic reticulum opens,
releasing Ga
2J
from the sarcoplasmic reticulum into the sarcoplasm. The
concentration of Ga
2J
in the sarcoplasm rises rapidly to 10&' mol12. The Ga
2J
&binding
sites on TpG in the thin filament are .uicly occupied by Ga
2J
. The TpGT%Ga
2J

interacts with Tp: and TpT to alter their interaction with tropomyosin. 6ccordingly,
tropomyosin moves out of the way or alters the conformation of A&actin so that the
myosin head&6,*&*
i
can interact with A&actin to start the contraction cycle.
$elaxation occurs when4
1. "arcoplasmic Ga
2J
falls below 10
&+
mol12 owing to its rese.uestration into the
sarcoplasmic reticulum by the Ga
2J
6T*aseN
2. TpGT%Ga
2J
loses its Ga
2J
N
#. Troponin, via its interaction with tropomyosin, inhibits further myosin head&
A&actin interactionN
%. :n the presence of 6T*, the myosin head detaches from A&actin.
Thus, Ga
2J
controls muscle contraction by an allosteric mechanism mediated in
muscle by TpG, Tp:, TpT, tropomyosin, and A&actin.
S$*3'/ $) /+/3&4 )$3 2*-'%/ <$3=
"ource of energy for all muscle is 6T*. :n .uiet conditions 1g of muscle
contains ' mmol of 6T* and in #&E times more another macroergic substances
creatine phosphate. This substance prevents the rapid depletion of 6T* by providing a
readily available high energy phosphate, which can be used to regenerate 6T* from
6,*. Greatine phosphate is formed from 6T* and creatine at times when the muscle
is relaxed and demands for 6T* are not so great. The enzyme catalyzing the
phosphorylation of creatine is creatine phosphoinase (G*S!, a muscle&specific
enzyme with clinical utility in the detection of acute or chronic diseases of muscle.
$eserves of 6T* and creatine phosphate can provide movement (action! during
)&10 se. 6fter this time oxidative phosphorylation of glucose, fatty acids and etone
bodies. This is enough for easy wor. 3ut in case of intensive wor (sprinter runner
for example! oxygen delivery does not cover necessity of muscle in energy. Thus
anaerobic glycolysis begins main way of 6T* resynthesis. Buscle glycogen and
glucose are decomposed to lactate.
,ifferent types of fiber have been detected in seletal muscle. Aor the sae of
simplicity, we shall consider 2 types4 type : (slow twitch, oxidative! and type :: (fast
twitch, glycolytic!. The type : fibers are red because they contain myoglobin and
mitochondriaN their metabolism is aerobic, and they maintain relatively sustained
contractions. The type :: fibers, lacing myoglobin and containing few mitochondria,
are white4 they derive their energy from anaerobic glycolysis and exhibit relatively
short durations of contraction.The proportion of these 2 types of fibers varies among
the muscles of the body, depending on function (eg, whether or not a muscle is
involved in sustained contraction, such as maintaining posture!. The proportion also
varies with trainingN for example, the number of type : fibers in certain leg muscles
increases in athletes training for marathons, whereas the number of type :: fibers
increases in sprinters.
!/'*%#(3#,4 $) '(31#(' 2*-'%/ 2/,(0$%#-2
Gardiac muscle also is striated. Rnlie seletal muscle, cardiac muscle exhibits
intrinsic rhytmicity, and individual myocytes communicate with each other because
of its syncytial nature. 6lso cardiac muscle has more mytochondria and less
myofibrils than seletal muscle. The T tubular system is more developed in cardiac
muscle, whereas the sarcoplasmic reticulum is less extensive and conse.uently the
intracellular supply of Ga
2J
for contaction is less. Gardiac muscle thus relies on
extracellular Ga
2J
for contraction. 6T* produced by oxidative phosphorilation
(aerobic glycolysis!. :n .uiet (rest! conditions 100g of heart tissue uses E&10 ml (
2

per minute (1' times more than other tissues!.
"ubstrates for oxidation are fatty acids, glucose, etone bodies, lactate and
pyruvate. 3ut fatty acids are the main substrate ()0&+0 8 of oxygen spent for their
oxidation!. ,uring wor utilization of glucose and lactate increases. 2actate is
produced by seletal muscle and transported by venous blood. 9eart has special
enzyme lactate dehydrogenase (2,51!, which transforms the lactate into pyruvate.
*yruvate used by mitochondria in oxidative decarboxylation and 6T* formed. 6lso
heart maintains the blood p9 due to this reaction.
Greatine phosphate also used in cardiac muscle and plays there double role
depo of energy and carrier of energy from mitochondria to myofibrils. 6T* produced
in mitochondria and gives the phosphate to creatine. Greatine phosphate pass through
mitochondria membrane toward cytoplasm and myofibrils. There creatine inase
removes phosphate from creatine phosphate to 6,*, which produced during
contraction. 7ew 6T* formed.
B#$'./2#-,34 $) ,3(#+#+&.
:n case of regular training in muscles activity of tissueHs dehydrogenases,
catalases and some other enzymes are increased. 6s a result in muscles synthesis of
glycogen, 6T*, creatin phosphate, specific muscles proteins is also increased. 6mount
of myosin increased. (xidative&reductive processes are increased and most of them
are aerobic (more effective!. "o, well trained muscles can perform more hard wor
longer and with less usage of energy than non&trained.
M*-'%/ )(,#@*/
Aati.ue of muscles during exercise is a phenomenon that almost everyone has
experienced. -hat is its causeV The primary cause is accumulation in muscle tissue
not of lactate (due to anaerobic glycolysis! but rather of protons. This fact has been
demonstrated by infusing lactate and observing that fati.ue does not necessarily
follow. :ncrease of protons (decreased p9! can affect the function of muscle in a
number of ways, including the following4
1. 2owering the O
max
of phosphofructoinase&1N
2. 2essening the release of Ga
2J
from the sarcoplasmic reticulumN
#. 2essening the activity of the actomyosin 6T*aseN
%. *ossibly by affecting the conformation of some of the muscle proteins
involved in contraction.
R/)/3/+'/-
1. Pohn -. "uttie. :ntroduction to 3iochemistry. 7ew >or4 9olt, $inehart and
-inston, :nc., 1//2.& #)% p.
2. Pohn Bc Burry, Bary ?. Gastellion. 5eneral, (rganic and 3iological
Ghemistry.& 7ew Persy4 *rentice 9all, 1//2.& +)% p.
#. $obert S. Burray, ,aryl S. 5ranner. 9arperHs illustrated 3iochemistry.
:ndia4 :nternational ?ducation, 200#.& )/# p.
*repared by :nna Srynytsa
$evised
6dopted at the Ghair "itting 1+.0).0'
Binutes W 12