Chapter 05

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Chapter 5

Membrane Structure, Synthesis and Transport

Key Concepts:

Membrane Structure Fluidity of Membranes Synthesis of Membrane Components Membrane Transport Transport Proteins

Exocytosis and Endocytosis

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Membrane: The fluid mosaic model Characteristics of the membrane: Fluidity: membrane is fluid (why?) Selective permeability: membrane is selectively permeable (why?) Components of the membrane Membrane transport: Passive transport: Passive diffusion & Facilitated diffusion Active transport: Primary active transport & Secondary active transport Transport of larger molecules: Exocytosis & Endocytosis: Endocytosis: Receptor mediated endocytosis, Pinocytosis & Phagocytosis Function and types of transport proteins: Channels Transporters Types of transporters: Uniporter, Symporter, Antiporter Specific examples of transport: Sodium Potassium Pump

Membrane Structure

The framework of the membrane is the phospholipid bilayer Phospholipids are amphipathic molecules

Hydrophobic (water-fearing) region faces in Hydrophilic (water-loving) region faces out

Membranes also contain proteins and carbohydrates

The two leaflets (halves of bilayer) are asymmetrical, with different amounts of each component
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Fluid-mosaic model

Membrane is considered a mosaic of lipid, protein, and carbohydrate molecules

Membrane resembles a fluid because lipids and proteins can move relative to each other within the membrane

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Extracellular environment
Carbohydrate Glycolipid Integral membrane protein Glycoprotein Phospholipid bilayer

Extracellular leaflet

HO

Polar

Cytosolic leaflet

Nonpolar

Peripheral membrane proteins

Cytosol

Cholesterol (found only in animal cells)

Polar

Proteins bound to membranes

Integral or intrinsic membrane proteins
Transmembrane

proteins

Region(s) are physically embedded in the hydrophobic portion of the phospholipid bilayer

Lipid-anchored

proteins

An amino acid of the protein is covalently attached to a lipid

Peripheral or extrinsic membrane proteins

Noncovalently

bound either to integral membrane proteins that project out from the membrane, or to polar head groups of phospholipids
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Extracellular environment
Transmembrane helix Transmembrane protein

Lipid

4
5

3 6

2 7 1

Lipidanchored protein

Peripheral membrane protein

Cytosol

Approximately 25% of All Genes Encode Transmembrane Proteins

Membranes are important medically as well as biologically Computer programs can be used to predict the number of transmembrane proteins

Estimated percentage of membrane proteins is substantial: 2030% of all genes may encode transmembrane proteins
This trend is found throughout all domains of life including archaea, bacteria, and eukaryotes Function of many genes is unknown study may provide better understanding and better treatments for disease

Transmission Electron Microscopy (TEM)

Biological sample is thin sectioned and stained with heavy-metal dyes Dye binds tightly to the polar head groups of phospholipids, but not to the fatty acyl chains

This makes membranes resemble railroad tracks

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Membrane bilayer
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Freeze Fracture Electron Microscopy (FFEM)

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specialized form of TEM used to analyze the interior of the phospholipid bilayer is frozen in liquid nitrogen and fractured with a knife

Direction of fracture

Sample

Transmembrane protein

Due

to the weakness of the central membrane, the leaflets separate into the P face (Protoplasmic face next to the cytosol) and the E face (Extracellular face)
provide significant detail about membrane protein form

P face exposed E face exposed

Cytosolic leaflet

P face

E face Extracellular leaflet E face

Can

P face

The McGraw-Hill Companies, Inc./Al Tesler, photographer

Fluidity of Membranes

Membranes are semifluid

Most lipids can rotate freely around their long axes and move laterally within the membrane leaflet But flip-flop of lipids from one leaflet to the opposite leaflet does not occur spontaneously Flippase requires ATP to transport lipids between leaflets
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Flippase Lateral movement

Flip-flop

Rotational movement
ATP (a) Spontaneous lipid movements ADP + Pi

(b) Lipid movement via flippase

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Lipid rafts

Certain lipids associate strongly with each other to form lipid rafts A group of lipids floats together as a unit within the larger sea of lipids in the membrane Composition of lipid raft is different than rest of membrane
High

concentration of cholesterol set of membrane proteins

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Unique

Factors affecting fluidity

Length of fatty acyl tails

Shorter

acyl tails are less likely to interact, which makes the membrane more fluid

Presence of double bonds

Double

bond creates a kink in the fatty acyl tail, making it more difficult for neighboring tails to interact and making the bilayer more fluid

Presence of cholesterol
Cholesterol Effects

tends to stabilize membranes

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vary depending on temperature

Experiments on lateral movement

Larry Frye and Michael Edidin experiment, 1970

Demonstrated the lateral movement of membrane proteins Mouse and human cells were fused Temperature treatment 0C or 37C Mouse membrane protein H-2 fluorescently labeled

Cells at 0C label stays on mouse side

Cells at 37C label moves over entire fused cell
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Add agents that cause mouse cell and human cell to fuse. Mouse cell H-2 mouse protein Human cell Fuse cells

Lower the temperature to 0C and add a fluorescently labeled antibody that recognizes the mouse H-2 protein in the plasma membrane. Observe with a fluorescence microscope. H-2 protein is unable to move laterally and remains on one side of the fused cell.

Incubate cell at 37C, then cool to 0C and add a fluorescently labeled antibody that recognizes the mouse H-2 protein in the plasma membrane. Observe with a fluorescence microscope. Due to lateral movement at 37C, the mouse H-2 protein is distributed throughout the fused cell surface.

Fluorescent dye H-2

Antibody

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Not all integral membrane proteins can move

Depending on the cell type, 1070% of membrane proteins may be restricted in their movement Integral membrane proteins may be bound to components of the cytoskeleton, which restricts the proteins from moving laterally Membrane proteins may be also attached to molecules that are outside the cell, such as the interconnected network of proteins that forms the extracellular matrix
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Fiber in the extracellular matrix Extracellular matrix

Plasma membrane
Linker protein Cytosol Cytoskeletal filament
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Glycosylation

Process of covalently attaching a carbohydrate to a protein or lipid

Glycolipid carbohydrate to lipid Glycoprotein carbohydrate to protein

Can serve as recognition signals for other cellular proteins

Often play a role in cell surface recognition Helps protect proteins from damage
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Membrane Transport

The plasma membrane is selectively permeable Allows the passage of some ions and molecules but not others This structure ensures that:

Essential molecules enter Metabolic intermediates remain Waste products exit

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Ways to move across membranes

Passive transport
Requires no input of energy down or with gradient
Passive

diffusion Diffusion of a solute through a membrane without transport protein diffusion Diffusion of a solute through a membrane with the aid of a transport protein

Facilitated

Active transport
Requires energy up or against gradient

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ATP ADP + Pi

(a) Diffusionpassive transport

(b) Facilitated diffusionpassive transport

(c) Active transport

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Phospholipid bilayer barrier

Barrier to hydrophilic molecules and ions due to hydrophobic interior

Rate of diffusion depends on chemistry of solute and its concentration Example: Diethylurea diffuses 50 times faster through the bilayer than urea, due to nonpolar ethyl groups
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O NH2 C Urea NH2 CH3 CH2 NH

O C NH CH2 CH3
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Diethylurea

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Artificial bilayer

Gases

CO2 N2 O2

High permeability

Very Ethanol small, uncharged molecules

Water Moderate permeability Urea

H2O

H2NCONH2

Low permeability

Sugars Polar organic molecules Ions Charged polar molecules and macromolecules Na+, K+, Mg2+, Ca2+, Cl Amino acids ATP Proteins Polysaccharides Nucleic acids (DNA and RNA)

Very low permeability

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Cells maintain gradients

Living cells maintain a relatively constant internal environment different from their external environment

Glucose

Transmembrane gradient

Concentration of a solute is higher on one side of a membrane than the other

Plasma membrane (a) Chemical gradient for glucosea higher glucose concentration outside the cell

+ +

+ +

Ion electrochemical gradient

+ + +

+ + + + + + + + + + + + +

Cl Na+ K+

Both an electrical gradient and chemical gradient

+ + +

Plasma membrane

(b) Electrochemical gradient for Na+more positive charges outside the cell and a higher Na+ concentration outside the cell

Tonicity

Isotonic
Equal

water and solute concentrations on either side of the membrane

Hypertonic
Solute

concentration is higher (and water concentration lower) on one side of the membrane

Hypotonic
Solute

concentration is lower (and water concentration higher) on one side of the membrane
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The solute concentration outside the cell is isotonic to the inside of the cell.

Solute

Cytosol (a) Outside isotonic

The solute concentration outside the cell is hypertonic to the inside of the cell.

(b) Outside hypertonic

The solute concentration outside the cell is hypotonic to the inside of the cell.

(c) Outside hypotonic

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Osmosis

Water diffuses through a membrane from an area with more water to an area with less water
If the solutes cannot move, water movement can make the cell shrink or swell as water leaves or enters the cell Osmotic pressure the tendency for water to move into any cell
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2% sucrose solution

1 liter of distilled water

1 liter of 10% sucrose solution

1 liter of 2% sucrose solution

aka: crenate

Hypertonic Conditions

Isotonic Conditions

Outside the cell Isotonic

Inside the cell

The solution and cell are isotonic

Hypertonic

The solution is hypertonic to the cell

Hypotonic

The solution is hypotonic to the cell

Osmosis in animal cells

aka crenation

Osmosis in plant cells

aka: plasmolysis

Which solution is hypertonic to the other?

the cell contents the environment

Transport proteins

Transport proteins enable biological membranes to be selectively permeable (will allow diffusion or not)
2 classes

Channels (porins)

Transporters

Channel Proteins

Form an open passageway, normally polar inside. i.e. Aquaporins

Osmosis in animal cells

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Animal cells must maintain a balance between extracellular and intracellular solute concentrations to maintain their size and shape Crenation shrinkage of a cell in a hypertonic solution Osmotic Lysis swelling and bursting of a cell in a hypotonic solution

Cells are initially in an isotonic solution. Red blood cell Cells maintain normal shape. Place in hypertonic solution. Place in hypotonic solution.

H2O
H2O

Cells undergo shrinkage (crenation) because water exits the cell.

Cells swell and may undergo osmotic lysis because water is taken into the cell.

(a) Osmosis in animal cells

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Osmosis in plant cells

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A cell wall prevents major changes in cell size

Turgor pressure pushes plasma membrane against cell wall

Cell is initially in an isotonic solution. Vacuole Cells maintain normal shape. Place in hypertonic solution. Place in hypotonic solution. Plant cell

H2O

H2O

Maintains shape and size

Plasmolysis plants wilting because water leaves plant cells

Volume inside the plasma membrane shrinks, and the membrane pulls away from the cell wall (plasmolysis) due to the exit of water. (b) Osmosis in plant cells

A small amount of water may enter the cell, but the cell wall prevents major expansion.

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Osmosis in freshwater protists

Freshwater protists like Paramecium have to survive in a strongly hypotonic environment

To prevent osmotic lysis, contractile vacuoles take up water and discharge it outside the cell Using vacuoles to remove excess water maintains a constant cell volume

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Filled contractile vacuole 60 m

Vacuole after expelling water 60 m

(all): Carolina Biological Supply/Visuals Unlimited

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Agre Discovered That Osmosis Occurs More Quickly in Cells with Transport Proteins That Allow the Facilitated Diffusion of Water

Water can passively diffuse across plasma membranes, but some cell types allow water to move across the membrane much faster than predicted
Peter Agre and colleagues first identified a protein that was abundant in red blood cells, bladder, and kidney cells Channel-forming Integral Membrane Protein, 28kDa (CHIP28) Unlike controls, frog oocytes that expressed CHIP28 swelled up and lysed when put in a hypotonic medium CHIP28 was renamed Aquaporin, since it forms a channel that allows water to pass through the membrane

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Experimental level

Conceptual level CHIP28 mRNA RNA polymerase

Add an enzyme (RNA polymerase) and nucleotides to a test tube that contains many copies of the CHIP28 gene. This results in the synthesis of many copies of CHIP28 mRNA.

Enzymes and nucleotides

CHIP28 DNA

Inject the CHIP28 mRNA into frog eggs (oocytes). Wait several hours to allow time for the mRNA to be translated into CHIP28 protein at the ER membrane and then moved via vesicles to the plasma membrane. Frog oocyte Nucleus Cytosol

CHIP28 mRNA

CHIP28 protein is inserted into the plasma membrane. CHIP28 protein Ribosome

Place oocytes into a hypotonic medium and observe under a light microscope. As a control, also place oocytes that have not been injected with CHIP28 mRNA into a hypotonic medium and observe by microscopy .

Control

THE DATA

Oocyte

Oocyte rupturing

35 minutes CHIP28 protein

Control

CHIP28

Control

CHIP28

Courtesy Dr. Peter Agre. From GM Preston, TP Carroll, WP Guggino, P Agre (1992), Appearance of water channels in Xenopus oocytes expressing red cell CHIP28 protein, Science, 256(5055):3857

Transport Proteins

Transport proteins are transmembrane proteins that provide a passageway for the movement of ions and hydrophilic molecules across membranes
Two classes based on type of movement
Channels Transporters

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Channels
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Form an open passageway for the direct diffusion of ions or molecules across the membrane Most are gated example: Aquaporins

When a channel is open, a solute directly diffuses through the channel to reach the other side of the membrane.
Gate opened Gate closed

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Transporters
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Also known as carriers

Hydrophilic pocket

Conformational change

Conformational change transports solute across membrane Principal pathway for uptake of organic molecules, such as sugars, amino acids, and nucleotides

Solute

For transport to occur, a solute binds in a hydrophilic pocket exposed on one side of the membrane. The transporter then undergoes a conformational change that switches the exposure of the pocket to the other side of the membrane, where the solute is then released.

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Transporter types
Uniporter

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A single solute moves in one direction.

Single molecule or ion

(a) Uniporter

Symporter

or cotransporter
Two or more ions or molecules transported in same direction

Two solutes move in the same direction.

(b) Symporter

Two solutes move in opposite directions.

Antiporter

Two or more ions or molecules transported in opposite directions

(c) Antiporter

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Question
A cell is placed in an hypertonic solution. Which way will the water move?
a. b.

Into the cell Out of the cell

c.

No net movement

Question
Gated channels which open when a chemical binds to it is
a. b. c.

Ligand gated channel Leakage channel Mechanically gated channel

d.
e.

Voltage gated channel

All can open in response to chemical binding

Question
What type of transport protein can move 2 or more different molecules in opposite directions?
a. b.

Uniporter Antiporter

c.
d. e.

Symporter
Multiporter Diporter

Active transport

Movement of a solute across a membrane against its gradient from a region of low concentration to higher concentration Energetically unfavorable and requires the input of energy Primary active transport uses a pump
Directly

uses energy to transport solute

Secondary active transport uses a different gradient

Uses

a pre-existing gradient to drive transport

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A pump actively exports H+ against a gradient.

Extracellular environment

A H+/sucrose symporter uses the H+ gradient to transport sucrose against a concentration gradient into the cell.

ATP

ADP + Pi
Sucrose Cytosol H+

(a) Primary active transport

(b) Secondary active transport

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ATP-driven ion pumps generate ion electrochemical gradients

Na+/K+-ATPase

Actively transports Na+ and K+ against their gradients using the energy from ATP hydrolysis 3 Na+ are exported for every 2 K+ imported into cell

Antiporter ions move in opposite directions Electrogenic pump exports one net positive (+) charge

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Nerve cell 1 3 Na+ bind from cytosol. ATP is hydrolyzed. ADP is released and phosphate (P) is covalently attached to the pump, switching it to the E2 conformation. Na+/K+-ATPase 3 Na+ Extracellular environment E2 2 3 Na+ are released outside of the cell. 3 2 K+ bind from outside of the cell.

Extracellular environment High [Na+] Low [K+]

E2
3 Na+

4 Phosphate (Pi) is released, and the pump switches to the E1 conformation. 2 K+ are released into cytosol. The process 2 K+ repeats.
E1

ADP + Pi

E1 ATP 2 K+ Cytosol ATP 3 Na+

Pi 2 K+ Cytosol ADP P

Low [Na+] High [K+] (a) Active transport by the Na+ / K+-ATPase

(b) Mechanism of pumping

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Exocytosis and Endocytosis

Used to transport large molecules such as proteins and polysaccharides

Exocytosis
Material

inside the cell packaged into vesicles and excreted into the extracellular medium

Endocytosis
Plasma

membrane invaginates (folds inward) to form a vesicle that brings substances into the cell Three types of endocytosis:

Receptor-mediated endocytosis Pinocytosis Phagocytosis

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Exocytosis
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Golgi apparatus

Cargo Vesicle

Cytosol

1 A vesicle loaded with cargo is formed as a protein coat wraps around it.

Protein

coat The vesicle is released from the Golgi, carrying cargo molecules.

Extracellular environment

3 The protein coat is shed. 4 The vesicle fuses with the plasma membrane and releases the cargo to the outside.
Plasma membrane

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Receptor-mediated endocytosis
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Cargo Invagination Coat protein

Cytosol

Cargo is released into the cytosol.

Receptor

Lysosome Extracellular environment

3 The protein coat is shed. 2 The vesicle is released in the cell.

The vesicle fuses with an internal organelle such as a lysosome.

Cargo binds to receptor and receptors aggregate. The receptors cause coat proteins to bind to the surrounding membrane. The plasma membrane invaginates as coat proteins cause a vesicle to form.

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NEXT QUIZ
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