CellBiology

AtifH.Khirelsied,B.Sc.,M.Sc.,Ph.D. Atif H. Khirelsied, B.Sc., M.Sc., Ph.D.

DepartmentofBiochemistry

FacultyofMedicine
InternationalUniversityofAfrica,Khartoum,Sudan

Thecell Thecell
Thecellisthebasic unitofbothstructure and Th ll i h b i i fb h d function in all living things inalllivingthings.

Animal and plant cell structure

Different Types of Cells DifferentTypesofCells

Therearetwomaingroupsofcells,prokaryotic andeukaryotic cells.

They differ not only in their appearance but also in Theydiffernotonlyintheirappearance butalsoin theirstructure,reproduction,andmetabolism. p

Prokaryoticcells Prokaryotic Prokaryotic cells

Prokaryoticorprocaryotic ,(fromtheGreekmeaning beforenuclei before nuclei beforenuclei). nuclei) Cellsthatareorganismsontothemselves,suchas g , bacteriacells. Theylackamembraneboundnucleus.

Eukaryoticcells Eukaryotic cells

DerivedfromtheGreek (good)and (karyon,"nut"or"kernel")

Cellsthathavemembraneboundnucleus.

Differencesbetweenprokaryoticand e e ces bet ee p o a yot c a d eukaryoticcells.

1. Prokaryotesareverysmall,aboutthesizeofa y y , mitochondrion (12mindiameterand10m long).. 2. Prokaryotesreproducebyasimplebinaryfission, comparedtocomplexreproductionprocessesin compared to complex reproduction processes in eukaryotes. y

Prokaryoticcellsdifferfromeukaryoticcells.
Property
DNA Size Si Organization

Eukaryotes
Linearstrandswithin membraneboundnucleus 5100m 5 100 Oftenmulticellular,some havecellwalls(no have cell walls (no peptidoglycan) Usuallyneedoxygento Usually need oxygen to exist Membranebound organelleslike mitochondria Plants,animals,protists, fungi

Prokaryotes
Singlecircleinnucleoid region 0.210m 0 2 10 Usuallysinglecelled, somehavepeptidoglycan some have peptidoglycan cellwalls Maynotneedoxygento May not need oxygen to exist Noorganelles,different ribosomes Bacteria,archaea
10

Metabolism Organelles

Examples

Eukaryoteshaveseveralmembranebound organelles.
1. 2. 2 3. 4. 5. 6. 7. 7 Theplasmamembrane. Thenucleus The nucleus Themitochondria Theendoplasmicreticulum g pp Golgiapparatus. Thelysosomes. Theribosomes nucleoproteinparticles,withno The ribosomes (nucleoprotein particles with no nucleoproteinparticles,withno membranesboundary)

Cellfractionation

Cellfractionation Cell fractionation

Itistheseparationofhomogeneoussets,usually It is the separation of homo eneo s sets s all g , p p organelles,fromapopulationofcells..

Hasthreeprincipalsteps:
1. Disruption(homogenization)ofcellsandliberation 1 Disruption (homogenization) of cells and liberation oforganelles. 2. Macrofiltration 2 Macro filtration 3. Purificationofcellcomponents.

Homogenization
Tissueishomogenizedinanisotonicbuffersolution. h d b ff l Thepopularprocedureistheuseof'PotterElvehjem homogeniser. g Other homogenization procedures include: grinding Otherhomogenizationproceduresinclude:grinding, mincing,chopping,pressurechanges,osmoticshock, freezethawing,andultrasoundhomogenization. freeze thawing and ultra sound homogenization

Homogenization
Homogenization is carried out in an isotonic solution Homogenizationiscarriedoutinanisotonicsolution, withapHbuffer,andaticecoldtemperatureto preventproteindamage. prevent protein damage Theseparatedorganellesarekepteithercold,isotonic orbuffered.

Purification
It is carried out by differential ultracentrifugation Itiscarriedoutbydifferentialultracentrifugation. Thesequentialincreaseinrelativecentrifugalforce (RCF)resultsinthesequentialseparationoforganelles accordingtotheirdensity. TheRCFisexpressedbyingunits,where1gisthe forceoftheearthsgravity. g y

DifferentialCentrifugationofLiverCellsHomogenate
The homogenate The Nuclear Supernatant p

Mitochondrial Supernatant

1. Centrifuge at 700 g for 10 min

2. Centrifuge at 20 000 g for 15 min

The Nuclear Pellet

Mitochondrial Pellet Centrifuge at 105 000 g for 60 min Microsomal Supernatant

Copy right Atif H Khirelsied 2006 .

Microsomal Pellet

Equilibriumgradientcentrifugation

Therelativecentrifugalforce(RCF) g ( )
Theforceexertedonaparticleinacentrifugeis expressedbythefollowingequation:

Where(rpm) istherevolutionsperminuteoftherotorand (r) isthedistance(incm)oftheparticlefromtheaxisof ( ) i th di t (i ) f th ti l f th i f rotation.

Thecellorganelles

TheplasmaMembrane The l Th plasmaMembrane M b

1. Theplasmamembrane(cellmembrane)ismade oftwolayersofamphipathiclipids.

2. Itconsistsofabilayer ofphospholipidsand 2 It consists of a bilayer of phospholipids and gy p glycolipids andanalmostequalamountof q proteins.

Schematicdiagramofaplasmamembraneshowingthemajor macromolecularcomponents.

Plasma membrane structure

5%

75%

20%

Theplasmamembrane The plasma membrane

3. Theproteinsareembedded insideofthecell membrane.

4. Thelipidcontentofthemembraneallowsthecell membranetospontaneouslyrepairitselfwhenitis torn.

Thefunctionoftheplasmamembrane
1. Itregulatestheentryandexitofsubstances g y (selectivepermeability). 2. Manymoleculescrossthecellmembraneby diffusion d diff i andosmosis,othersbytransport i th b t t systems. 1. Differenttransportsystems existintheplasma p y p membrane.

Plasmamembranetransportsystems Plasmamembranetransportsystems

2.

TheNucleus The Nucleus

1. Isadoublemembraneboundedorganelle,(only ineukaryotes) i k )

2. Thenucleusisthecellcontrolcenter,Itstoresthe geneticinformation.

TheNucleus
3. Thegeneticmaterial,theDNAiscomplexed with 3 Th ti t i l th DNA i l d ith histones andorganizedintochromatin. g

4. Duringdivision,thechromatinsarecondensedto formhighlyorganizedchromosomes.

TheNucleus The Nucleus

5. Thenucleusisgenerallyfoundatthecenterofthe cell. ll

6. Besidesthegeneticmaterial,mostnucleialso containnucleoli(singlenucleolus)

TheNucleus The Nucleus

The nucleus
Nucleolus.

Nuclearmembrane
Micrographofacellnucleus showingthedarklystained nucleolus.

TheMitochondrion

TheMitochondrionfunction The Mitochondrion function

Mitochondriahavemanydifferentfunctions. Including;
1. Energytransformation andATPgeneration. 2. Metabolicprocesses suchasheme synthesis,urea formation,etc. 3. HasitsownDNA whichissimilartobacterialDNA. 3 Has its own DNA which is similar to bacterial DNA

TheEndoplasmicReticulum(ER) The Endoplasmic Reticulum (ER)

Itisanextensivenetworkoffinebranchedacisternae (saclikestructures)heldbythecytoskeleton. The ER membrane divides the cytoplasm into two TheERmembranedividesthecytoplasmintotwo phases; Intra cisternal (luminal) phase Intracisternal (luminal)phase Hyaloplasmic phase(cytoplasm)

TheroughandsmoothER. h h d h
TheERassociatedwithribosomes isknownasrough ER,theonelackingribosomes i th ER th l ki ib isthesmoothER. th ER

TheRERisinvolvedinthesynthesisofproteins,while SERis involvedinthesynthesisoflipids,andseveral otherfunctionsasregulationofcalciumconcentration th f ti l ti f l i t ti anddetoxificationofdrugs.

EndoplasmicReticulum

Ribosomes
They are nucleoproteins particles Theyarenucleoproteins particles.

TheymayeitherexistassociatedtotheER,orfound assembledinapolysomes.

Ribosomes areresponsibleforproteinsynthesis.

Ribosomestructure Ribosome structure

Theribosomeconsistsofalargeandsmall subunits. b i

Half of the ribosome weight is RNA, the remaining HalfoftheribosomeweightisRNA,theremaining arevariousproteins.

Ribosomestructure

Ribosomestructure

Golgiapparatus(complex) Golgi apparatus (complex)

Itconsistsofasetofsmoothcisternae whichare stackedtogetherinparallelrows. k d h i ll l

Has a variety of functions, including sorting and Hasavarietyoffunctions,includingsortingand glycosylation andsecretionofproteins..

TheLysosome The Lysosome

Theyaresinglemembranebound,vesicular structures. structures Generallysmallerthanmitochondria. Containsvarioushydrolyticenzymes(hydrolases).

Thelysosome The The lysosome function

Responsible for hydrolytic digestion of intracellular Responsibleforhydrolyticdigestionofintracellular molecules. Containsover40differentprotease,nuclease,and phopholipase enzymes. Deficiencyofcertainhydrolases causesseriousdiseases. e.g., eg TaySachs disease(deficiencyofhexosaminidase A).
NiemannPick disease(deficiencyofsphingomyelinase) disease

Thecytoskeleton The cytoskeleton

1. Eukaryoticcellshaveawidevarietyofdistinct shapesandinternalorganizations. h di t l i ti

3. Thechangeofshapeandmovement isachievedby anetworkaproteinfilaments knownasthe cytoskeleton.

Cytoskeleton

TheCytoskeleton y

Thefunctionofthecytoskeleton The function of the cytoskeleton

Thetwomostimportantproteinfilamentsare h fl calledtheactin filaments andthemicrotubules. The actin is responsible for contraction (like in Theactin isresponsibleforcontraction (likein muscles). Themicrotubulesareresponsibleforstructural strength.