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BIOCHEMISTRY

Biological Membrane and Transport | Dr. Laygo | (10/7/19)

OUTLINE
Transcriber 1: I. BIOLOGICAL MEMBRANES
I. BIOLOGICAL MEMBRANES  Defined as external boundaries of cells.
II. BIOLOGICAL ACTIVITIES OF BIOLOGICAL MEMBRANES
 Regulate molecular traffic across the boundary.
III. MACROMOLECULAR CONSTITUENTS OF MEMBRANES
A. Lipids  Divide the internal space of eukaryotic cells into
B. Proteins discrete compartments to segregate processes and
C. Carbohydrates components.
IV. COMPOSITION AND ARCHITECTURE OF MEMBRANES
 Organize complex reaction sequences that are
V. COMMON FEATURES OF MEMBRANE
VI. FUNCTIONS OF PROTEINS IN MEMBRANES central to both biological energy conservation and
VII. THREE TYPES OF MEMBRANE PROTEINS DIFFER IN THEIR cell-to-cell communication.
ASSOCIATION WITH MEMBRANES
VIII. CHEMICAL FORCES THAT STABILIZE LIPID BILAYERS
IX. CLINICAL CORRELATION: LIPOSOMES AS VEHICLES FOR
DRUG DELIVERY
X. MEMBRANE PROTEINS
A. Integral proteins
A.1. Alpha-helix
A.2. Beta-barrel

XI. MEMBRANE DYNAMICS


A. Acyl groups in the bilayer interior
B. Transbilayer movement
B.1. Flipapses
B.2. Floppases
B.3. Scramblases
C. Study of membrane dynamics: FRAP
D. Membrane rafts, membrane fusion, and
neurotransmitter release
E. Integral proteins: Surface adhesion, signaling, and other
cellular processes
II. BIOLOGICAL ACTIVITIES OF BIOLOGICAL MEMBRANES
XII. SOLUTE TRANSPORT ACROSS MEMBRANES  FLEXIBILITY- permits shape changes that accompany
A. Simple diffusion cell growth and movement.
B. Passive transport facilitated by membrane proteins  ABILITY TO BREAK AND RESEAL- two membranes can
A.2. Group of transporters based on structures fuse (exocytosis); a single membrane -enclosed
A.3. Example of transporters compartment can undergo fission to yield two sealed
i. Glucose transporters (GLUT’s) compartments (endocytosis or cell division)
ii. Chloride-bicarbonate exchanger  SELECTIVELY PERMEABLE- retain certain compounds
C. Active transport and ions within cells or specific cellular
B.1. Primary and secondary active transport compartments, while excluding others.
B.2. Four general types of transport ATPase
i. P-type
ii. F-type
iii. V-type
iv. ABC transporters
B.3. Ion gradients and secondary active transport
(p.43,45)
D. Other channels
i. Ion-selective
ii. Ligand-gated
iii. Voltage-gated

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Biological Membrane and Transport | Dr. Laygo | (10/7/19)

III. MACROMOLECULAR CONSTITUENTS OF MEMBRANES plane of the membrane.

A. Lipids
 Cholesterol
 Sphingolipids
◦ sphingomyelin (SP)
◦ gangliosides
 Glyceryl Phospholipids
◦ Phosphatidylcholine (PC)
◦ Phosphatidylthanolamine (PE)
◦ Phosphatidylglycerol (PG)
◦ Phosphatidylserine (PS)
◦ Phosphatidylinosotol (PI)
◦ Cardiolipin (CL)

B. Proteins
 Classified as integral and peripheral  A Lipid bilayer is the basic structural element of
membranes.
 Three (3) types of lipid aggregate.
C. Carbohydrates  MICELLES
 In the form of glycoprotein and glycolipid, never - forms in the solution of amphiphatic
free. molecules that have larger head than
tail (Fatty acid, dodecyl sulfate)
IV. COMPOSITION AND ARCHITECTURE OF MEMBRANES - Each miscelle has from a few dozen to
few thousand lipid molecules.
- Aggregation occurs when the
concentration of molecules is higher
than a certain threshold.
 BILAYER
- Has two (2) lipid monolayer (leaflets)
forms a 2D sheet.
 VESICLE (LYPOSOME)
- small bilayers will spontaneously seal
into spherical vesicles
- Vesicle membranes can contain
artificially inserted proteins
- The central aqueous cavity can enclose
and dissolve molecules
 All biological membranes share some fundamental - Useful Carriers of molecules (ie. Drugs)
properties - Vesicles fuse readily with cell
 Fluid mosaic model for the structure of membranes or with each other.
biological membranes. -
 Phospholipids and sterols form a V. COMMON FEATURES OF MEMBRANE
bilayer in which the nonpolar regions of  Sheet-like flexible structure, 30-100Å (3-10mm) thick
the lipid molecules in each layer face  Main structure composed of two leaflets of lipids
outward, interacting with the aqueous (bilayer), except for archaebacteria: which has a
phase on either side. monolayer of bifunctional lipid.
 Proteins are embedded in this bilayer  Forms spontaneously in aquaous solution and are
sheet, held by hydrophobic interactions stabilized by non-covalent forces (esp. hydrophobic
between the membrane lipids and effect)
hydrophobic domains in the protein  Protein molecules span the lipid bilayer
 Membrane is fluid (lipid and protein  Assymetric
molecules free to move laterally in the ◦ Some lipids are found preferably “inside”

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BIOCHEMISTRY
Biological Membrane and Transport | Dr. Laygo | (10/7/19)

◦ Some lipids are found preferably “outside”  found in both the cytosol and in association with
◦ Carbohydrate moieties are always outside the membranes.
cell
◦ Electrically polarized (inside -60mV) VIII. CHEMICAL FORCES THAT STABILIZE LIPID BILAYERS
 Fluid structures: 2-dimensional solution of oriented
lipids. A. Hydrophobic interactions
 Hydrophobic interactions are the PRIMARY force.
These occur between the extensive hydrophobic lipid
tails that are stacked in the sheet.
B. Van der waals
 Van Der Waals attractive forces between the
hydrocarbon tails favor their close packing.

B. Electrostatic interactions
 Electrostatic interactions lead to hydrogen bond
formation between the polar head groups and water
molecules in the solution.

Note: THEREFORE, THE SAME CHEMICAL FORCES THAT


STABILIZE PROTEIN STRUCTURES SATBILIZE LIPID BILAYERS

IX. CLINICAL CORRELATION: LIPOSOMES AS VEHICLES FOR


DRUG DELIVERY
1. The propensity of phospholipids to form bilayers has
been exploited to create an important clinical tool :
THE LIPOSOME
VI. FUNCTIONS OF PROTEINS IN MEMBRANES
2. When suitable phospholipids are sonicated (agitated
 RECEPTORS: detecting signal from outside by high frequency sound waves) in aqueous solution,
 Light (opsonin) they form ~50nm lipid vesicles, also called
 Hormones (insulin receptors) liposomes.
 Neurotransmitters (acetylcholine receptors)
 Pheromones (taste and smell receptors)
 Channels, gates, pumps
 Nutrients (maltoporin)
 Ions (K-channels)
 Neurotransmitters (serotonin reuptake protein)
 Enzymes
 Lipid biosynthesis (some acyltransferases)
 ATP synthesis (F0F1ATPase/ATP synthase)

VII. THREE TYPES OF MEMBRANE PROTEINS DIFFER IN THEIR


ASSOCIATION WITH MEMBRANES
 INTEGRAL PROTEIN
 very firmly associated with the lipid bilayer, and
are removable only by agents that interfere with
hydrophobic interactions. LIPOSOMES CAN BE ENGINEERED TO CONTAIN HYDROPHILIC
 PERIPHERAL MEMBRANE PROTEINS MOLECULES IN THEIR AQUEOUS CENTER
 associate with the membrane through
electrostatic interactions and hydrogen bonding
with the hydrophilic domains of integral 1. Drugs or DNA for gene therapy may be incorporated
proteins and with the polar head groups of into a liposome, which can then be injected into a
membrane lipids. patient.
 AMPHITROPIC PROTEINS 2. Liposome fuse with the plasma membrane, and

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BIOCHEMISTRY
Biological Membrane and Transport | Dr. Laygo | (10/7/19)

deliver their contents directly into cells, bypassing


both the circulation and the digestive system.
3. In the future, targeting signals may be incorporated
into the liposomes to allow more selective drug
delivery.

X. MEMBRANE PROTEINS  Example of a one transmembrane segment with


globular domains on either end.
 Transmembrane segment is alpha helical and
consists of 19 hydrophobic amino acids
 Extracellular portion contains oligosaccharides (and
these constitute the ABO and MN blood group
determinants)
 held in the membrane by hydrophobic interactions
with lipids

A. Peripheral proteins

PERIPHERAL MEMBRANE PROTEINS


- can be relieved by mild conditions, such as : change in pH or
ionic strength, add urea. If covalently linked to membrane
lipid, the use of phospilipase C to cleave the linkages.

B. Integral proteins
 The firm attachment of integral proteins to
- can be relieved by the use of detergent. membranes is the result of hydrophobic interactions
between membrane lipids and hydrophobic
SOME MEMBRANE PROTEINS SPAN THE LIPID BILAYER domains.
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BIOCHEMISTRY
Biological Membrane and Transport | Dr. Laygo | (10/7/19)

 Integral proteins can be classified into six (6)  10-20% of all membrane proteins: integral
categories/ membrane proteins
 Type I and II : one transmembrane helix.
 Type III: multi-transmembrane helices in a single  Hydropathy Index: the relative polarity of each
polypeptide chain. amino acid side chain determined by measuring the
 Type IV: transmembrane domains from different free energy change accompanying the movement of
polypeptides assemble to form a channel an amino acid from a hydrophobic solvent to water.
 Type V: held to the bilayer through covalently
linked lipids.  Hydropathy Plots are used to search for 20-amino
 Type VI: contains both transmembrane helices acid streches of hydrophobic amino acids in the
and GPI-anchor. primary sequence of a protein for which a crystal
structure is not available.
NOTE: SINGLE PASS and MULTIPLE PASS proteins.  Putative hydrophobic transmembrane α-helices
have been identified this way in many
Example: Bacteriorhodopsin membrane proteins.
 A membrane spanning protein
 Light-driven proton pump  Hydropathy plots alone are not conclusive.
 Seven transmembrane domains  PROTEIN TOPOLOGY studies are used to test the
 Multi-pass transmembrane distribution of protein domains
predicted by hydropathy plots.
 If a hydropathy plot indicates ONE 20-amino acid
hydrophobic stretch (1 putative transmembrane α-
helix), topology studies are expected to confirm
location of N & C termini on opposite sides of the
membrane.
 If TWO transmembrane α-helices are predicted, N &
C termini should be on the same side. The segment
between the α-helices should be on the other side.

INTEGRAL PROTEIN STRUCTURE


THE TOPOLOGY OF AN INTEGRAL MEMBRANE PROETIN CAN
BE PREDICTED FROM ITS SEQUENCE

membrane

B.1. Alpha helix

 Unbroken sequences of more than 20


hydrophobic residues

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Biological Membrane and Transport | Dr. Laygo | (10/7/19)

-helix
R-groups in magenta

 A membrane-spanning α-helix is the most common


structural motif found in integral proteins.
 In an α-helix, amino acid R-groups protrude out
from the helically coilded polypeptide backbone. The
largely hydrophobic R-groups of a membrane-
spanning α-helix contact the hydrophobic  Lysine & arginine are often at the lipid/water
membrane core, while the more polar peptide interface, with the positively charged groups at the
backbone is buried. ends of their aliphatic side chains extending toward
the polar membrane surface.
 COLORS C, N, O, R-group (H atoms not shown)

 Particular AMINO ACIDS tend to occur at different


positions relative to the surface or interior of the
bilayer in transmembrane segments of integral
proteins.

 Residues with ALIPHATIC side-chains (leucine,


isioleucine, alanine, valine) predominate in the
middle of the bilayer.

alanine (Ala, A) isoleucine (Ile, I) leucine (Leu, L) valine (Val, V)

H H H H

H3N+ C COO H3N+ C COO H3N+ C COO H3N+ C COO

CH3 CH CH3 CH2 CH CH3  CYTOCHROME OXIDASE is an integral protein whose


CH2 CH CH3 CH3
intramembrane domains are mainly transmembrane
α-helices.
CHaliphatic
amino acids: non-polar 3 CH3
R-groups

 Tyrosine and tryptophan are common near the


membrane surface.

membrane

Cytochrome oxidase dimer (PDB file 1OCC)

B.2. Beta-barrel
 It has been suggested that the polar character of the
tryptophan amide group and the tyrosine hydroxyl,
along with their hydrophobic ring structures, suit
them for localization at the polar/apolar interface.

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BIOCHEMISTRY
Biological Membrane and Transport | Dr. Laygo | (10/7/19)

Porin Monomer

PDB 1AOS
(Above is shown one channel of a trmeric porin complex)

 Porin β-barrel
 While transmembrane α-helices are the most
common structural motif for integral proteins, a
family of bacterial outer envelope channel proteins
called porins have instead β-barrelstructures. XI. MEMBRANE DYNAMICS
 A β-barrel is a β-sheet rolled up to form a cylindrical
pore. A. Acyl groups in the bilayer interior
 Although the lipid bi-layer is stable, its individual
phospholipid and sterol molecules have much
freedom of motion.
 The structure and flexibility of the lipid bi-layer
depends on the kinds of lipids present, and change in
temperature.
 Below normal physiological temperatures, the lipids
 polar R group,  non-polar R group in a bi-layer form a semisolid gel phase.

 In a b-sheet, amino acid R-groups alternately point


above & below the sheet. Much of porin primary
structure consists of alternating polar & non-polar
amino acids.
 Polar residues face the aqueous lumen.
 Non-polar residues are in contact with membrane
lipids.

MEMBRANE PROTEINS WITH BETA-BARREL STRUCTURE:

B. Transbilayer Movement

Transbilayer movement of lipids requires a catalyst. Flip-flop


of lipids is negligible. Polar headgroup doesn’t want to enter
membrane.

B.1 Flipapses:

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Biological Membrane and Transport | Dr. Laygo | (10/7/19)

 Catalyzes translocation of the amino phospholipid;


phosphatidylethanolamine and phosphatidylserine
from the extracellular to the cytosolic leaflet of the
plasma membrane.

 translocates aminophospolipids specifically


phosphotidyl serine and phosphotidylethanoamoine
from extracellular leaflet to cytosolic leaflet (Outside
to Inside)
o Creates asymmetric distribution of
Phospho-ser and –ethanol on inside the
membrane; sphinolipids and phosphololine
outside the cell uses ATP
o Important because having phosphoser
outside is to trigger apoctosis.

B.2 Floppase:
 Moves plasma membrane phospholipids from the
cytosolic to the extracellular membrane.
 translocate phospholipids from cytosolic leaflet to
extracellular leaflet (inside to outside)
o Also ATP dependent

B.3 Scramblases:
 Proteins that move any membrane phospholipid C. Study of membrane dynamics: FRAP
across the bi-layer down its concentration gradient.
 Move any phospholipid across bilayer to follow  Fluorescence Recovery after Photobleaching allows
concentration gradient to monitor lateral lipid diffusion by monitoring the
o Does not need ATP rate of fluorescence return
o Faster when Ca2+ is present  From the rate of return of lipids, the diffusion
 *Phosphatidylinositol transfer protein are used in coefficient of a lipid in the leaflet can be determine
lipid signaling and cell trafficking

Lipids and proteins diffuse laterally; lateral diffusion is fast.


Can take only about a second for lipid to circumnavigate the
cell so composition of inner or outer surface quickly
homogenizes self.

D. Membrane rafts, membrane fusion, and neurotransmitter


release

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Biological Membrane and Transport | Dr. Laygo | (10/7/19)

D.1 Membrane Rafts  Ability of membrane to fuse without losing


 Lipid distribution in a single leaflet is not random continuity is very important because cellular
 Some regions contain clusters of glycosphingolipids membrane, from nucleus, ER, Golgi, and various
with longer than usual tails small vesicles are constantly reorganizing.
 These regions are more ordered and contain specific  Also included are exocytosis, endocytosis, cell
doubly or triply acylated proteins division, fusion of egg and sperm.
 Allow segregation of proteins in the membrane  Most of the following start with an increase of
curvature of a local area in the membrane.

Fusion of membranes requires the following steps:


1. Membrane recognizes each other
2. Surface becomes closely opposed – water is
removed from the surface
3. Local bilayer structure breaks down and outer
leaflets fuse
4. Bi-layers fuse

D.3 Neurotransmitter Release

Lipids can move in and out of the raft cell and its
surroundings. Depending on the type of cell, rafts make up to
50% of surface of the cell and may serve as “Glue” for the
membrane proteins that have to associate for its activity.

D.2 Membrane Fusion


 Membrane can fuse with each other without losing
continuity
 Fusion can be spontaneous or protein – mediated
 Examples of protein – mediated fusion:
o Entry of influenza virus into the host cell
o Release of neurotransmitter at nerve
synapse

Snare proteins are used when vesicle containing


neurotransmitter fuses with plasma membrane to release
transmitter into synapse:
 V-SNARE – found on cytoplasmic side of vesicle
 T-SNARE – found on target side of plasma membrane
 NSF and SNAP25 are also involved

*Botulism – Clostridium botulinum toxin specifically cleaves


SNARE protein – prevents neurotransmission and organisms
die.

E. Integral proteins: Surface adhesion, signaling, and other


cellular processes
 Integrins are surface adhesion proteins that mediate
a cell’s interaction with the extracellular matrix and
with other cells.
o Anchored to membrane by a single
transmembrane helix / subunit Alpha-Beta
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Biological Membrane and Transport | Dr. Laygo | (10/7/19)

Dimer site for extracellular proteins to


attach
o Different combinations of alpha or beta
proteins create a wide range of binding sites
o Large extracellular side of alpha and beta
protein are specific binding site for
extracellular proteins like collagen or
fibronectin
o Also used as receptors and signal
transducers.
 Other plasma proteins involved in surface adhesion
are cadherins, undergo hemophilic interactions with
identical cadherins in an adjacent cell. (Identical)
 Selectins, have extracellular domains that, in the
presence of Ca2+, bind specific polysaccharides on
the surface of an adjacent cell. (Identical)
o Part of blood clotting
 The direction of a charged solute tends to move
across a membrane depends on:
XII. SOLUTE TRANSPORT ACROSS MEMBRANE
(1) chemical gradient
- the difference in solute concentration
(2) electrical gradient across the membrane
- called electrochemical gradient or
electrochemical potential)

Electrochemical gradient
combination of both concentration gradient
and membrane potential

B. Passive transport facilitated by membrane proteins

 Energy changes accompanying passage of a


hydrophilic solute through the lipid bilayer of a
biological membrane

 Passive diffusion of polar molecules involves


desolvation and thus has a high activation barrier

A. Simple diffusion

 Happens when two aqueous compartments containing


unequal concentrations of a soluble compound or ion
are separated by a permeable divider (membrane)
 The solute moves from the region of higher
concentration, through the membrane, to the region of
lower concentration, until the two compartments have
equal solute concentrations

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Biological Membrane and Transport | Dr. Laygo | (10/7/19)

B.1. Group of transporters based on structures

B.2. Example of transporters


Table 1. The Transporter Classification (TC) System
*highlighted in orange were given emphasis
1.A. α-helix type channels
1.A.1. Voltage-gated ion channel VIC superfamily
Voltage-gated K+ channel
1.A.3 Ryanodine/IP3 receptor Ca2+ channel
1.A.8 Major intrinsic protein family
Aquaporins
1.A.9 Ligand-gated ion channel (LIC) of neurotransmitter
receptors
Notes from the graph (Dr. Laygo): Acetylcholine receptor/channel
 The graph can be compared to uncatalyzed and 1.B. β-barrel porins
enzyme-catalyzed reaction 1.B.1 General bacterial porin (GBP) family
 Red graph – similar to uncatalyzed reaction 1.C. Pore-forming toxins
 Blue graph – similar to enzyme-catalyzed reaction 1.C.7 Diphtheria toxin family
due to proteins that will facilitate the transport; there 1.C.18 Mellitin family (bee venoms)
is lowering in the energy of activation 2.A. Porters: uniporters, symporters, and antiporters
*this is similar to enzymes whereby the substrate is 2.A.1 Major facilitator superfamily (MFS)
converted to transition state (there is a Lactose transporter/ permease of E. coli
contemplation whether a bond will be broken or 2.A.1.1 Sugar porter family
formed) GLUT1 glucose transporter of erythrocyte
 The energy of activation will be involved in 2.A.1.9 P1-H+ symporter
desolvation or removal of water molecules thea
2.A.12 ATP-ADP antiporter (AAA) family
surround the polar solute (encircle in blue in the
2.A.13 C4-dicarboxylate uptake (Dcu) family
picture); +
2.A.21 Solute-Na symporter (SSS) family
Otherwise, if the water molecules will not be +
Na - glucose symporter in epithelial cells
removed, it will interact with the binding of the
2.A.73 HCO3—Cl- transporters
substrate/solure to the active site.
 Desolvation exposes the groups of the HCO3—Cl- antiporter
substrate/solute that will interact with the amino 2.B. Nonribosomally synthesized porters
acid R groups that are present within the channel 2.B.1 Valinomycin carrier family
Valinomycin
3.A. Diphosphate bond hydrolysis-driven transporters
(use PPi, not ATP)
3.A.1 ATP-binding cassette (ABC) superfamily
CFTR Cl- channel; multidrug transporter MDR1
3.A.2 H+- or Na+- translocating F-type, V-type, A-type
ATPase superfamily
FoF1 ATPase proton pump; VoV1 ATPase; AoA1
ATPase
3.A.3 P-type ATPase superfamily
Na+K+ ATPase antiporter; SERCA Ca2+ pump
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Additional Notes from Dr. Laygo (recording):

Ryanodine/IP3 receptor Ca2+ channel


 Found in muscles

ATP-ADP antiporter (AAA) family


 Important in electron transport chain

Na+-glucose symporter in epithelial cells


 Found in gastrointestinal cells or kidney cells

Valinomycin
 Inhibitor of ETC
 Kind of ion channel, especially for potassium

FoF1 ATPase proton pump


 Found in electron transport chain

Na+K+ ATPase antiporter


 Found in muscles

i. Glucose transporters of eythrocytes


Kinetics of glucose transport into erythrocytes
 Mediates passive transport

GLUT 1
 Some are found in the brain but GLUT 3 is the major
GLUT in the brain

Proposed structure of GLUT1

Graph on the left


Michelis-Menten graph

Equation:

Notes on the structure:


 Multi-pass; has an alpha-helical structure
 Both amino- and carboxy- groups are found inside
the cell
 Hydrophpobic proteins make up the transmembrane  A rectangular hyperbola
domain  At the plateau point (saturation point), no amount of
substrate concentration can be associated with an
Interior of GLUT1

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increase in the velocity of the reaction; all of the  Glucose is binded from the outside
active site are occupied by substrate  GLUT 1 undergoes a conformational change
 Values that can be determined from the graph are  It opens to the inside allowing glucose to get in
Vmax and Km (Km is the value at ½ Vmax)
Different types of GLUT
On a scale of 1 to 10, how would you rate your pain?
–Vmax

Graph on the right:


Lineweaver-Burke plot
GLUT 4
 Reciprocal of the equation by Michelis and Menten
 For the regulation of glucose of insulin into a
 Follows the slope-intercept plot (y=mx+b) myocyte
 Values that can be determined from the graph are
Km (from manipulation of the x intercept and Vmax
(from the manipulation of the y-intercept)

Lineweaver Burke equation from Michelis-Menten equation:

Receptor tyrosine kinase


- Receptor for insulin

Model glucose transport into erythrocytes by GLUT1 Suggested additional reading by Doc:
Action of insulin from Harper’s under hormones

ii. HCO3—Cl- antiporter

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Use this space to congratulate yourself for a job well done


(Charz nagulo lang talaga format di ko alam bat may space


dyan hahahaha)

THREE GENERAL CLASSES OF TRANSPORT SYSTEMS


Notes from recording:

HCO3—Cl- antiporter
 Found in red blood cells

*cell will generate CO2 as product of metabolism


 CO2 will go to RBC and will react with waterm
catalyzed by carbonic anhydrase to from
carbonic acid  carbonic acid will dissociate into
bicarbonate ion and protons (H+)  protons will
be bound by hemoglobin  hemoglobin serves
as a good buffer (hemoglobin binds the protons
while bicarbonate ions will get out of the RBC in C. Active transport
exchange for chloride ions – chloride shift)  Movement against electrochemical gradient
 Involves hydrolysis of ATP
*therefore most of the CO2 is not found in the
RBC but in the plasma in the form bicarbonate C.1. Primary and Secondary Active Transport
ion

*CO2 in the plasma  reach the lungs 


chloride shift again – bicarbonate from the
plasma will get inside the RBC in exchange for
chloride ions

*in the lungs O2 concentration > CO2, oxygen will


now be binded to hemoglobin, releasing the
protons; The protons will now bind again to
bicarbonate ions as catalyzed by carbonic
anhydrase to reform carbonic acid; in the lungs,
carbonic acid will spontaneously dissociate to *X solute
CO2 and H2O; CO2 will be expired

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Primary active transport From recording:


- the energy released by ATP hydrolysis drives
solute movement against an electrochemical In the gastrointestinal tract, solute X is
gradient represented by Na+ ions and S can be glucose
or amino acid as a result of final digestion of
Secondary active transport carbohydrate and proteins in the small
- A gradient of ion X (often Na+) has been intestine;
established by primary active transport.
Movement of X down its electrochemical Too much sodium inside the cell is not good
gradient now provides the energy to drive because it will cause the cell to burst.
cotransport of a second solute (S) against its Sodium that was taken in should be expelled
electrochemical gradient. and that is against the concentration
gradient; this is done in expense of an ATP
Symport example: Na+-glucose symporters (Na+-K+ ATPase pump)
 in the apical plasma membrane take up glucose from
the intestine in a process driven by the downhill flow
of Na+ Other cotransport systems:

Antiport example: lactose transporter (lactose permease) C.2. Four general types of transport ATPase

 The lactose transporter (lactose permease) i. P-type


of E. coli is the well-studied prototype for protondriven  Undergo Phosphorylation during their catalytic
cotransporters cycles
 The family of active transporters called P-type
ATPases are cation transporters that are reversibly
phosphorylated by ATP as part of the transport
cycle.
 In animal tissue, the Na+K+ATPase and Ca2+ ATPase
are ubiquitous P-type ATPase that maintain the
differences in the ionic composition of cytosol and
extracellular medium.
 The plasma membrane Ca2+ pump moves calcium
ions out of the cell, and another P-type pump in the
endoplasmic
 reticulum moves Ca2+ into the ER lumen.
 The sarcoplasmic and endoplasmic reticulum
calcium (SERCA) pumps are P-type ATPases closely
related in structure and mechanism.

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 A variation on this basic mechanism is seen in the


Na+K+ATPase of the plasma membrane, discovered
by Jens Skou in 1957.

ATP-drive Ca2+ pumps (P-type ATPase) maintain a low conc.


of calcium in the cytosol

 Calcium ions are pumped out of the cytosol by


plasma membrane Ca2+ pump (a P-type ATPase) to
maintain low conc. of calcium in the cytosol
 Calcium ions are pumped in to the lumen by
endoplasmic reticulum and sacoplasmic reticulum
Ca2+ pump (SERCA).
 SERCA contains a single polypeptide (Mr ~ 100,000)
and cycles among conformations (see Fig. 11-36). ii. F-type
 Two calcium ions bind to a transmembrane domain. - active transporters catalyze the uphill
Phosphorylation on Asp351 mediates transmembrane passage of protons driven by
conformational change and controls calcium ATP hydrolysis.
release/binding

iii. ABC Transporter


 ABC transporters constitute a large family
of ATP-dependent transporters that pump amino acids,
peptides, proteins, metal ions, various lipids, bile salts,
and many hydrophobic compounds.
 One ABC transporter in humans, the multi-drug
transporter (MDR1), is responsible for the striking
resistance of certain tumors to some generally effective
antitumor drugs.

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D. Channels

Ion-selective channels
 Ion channels, together with ion
pumps such as the Na+ K+ ATPase, determine a plasma
membrane’s permeability to specific ions and regulate the
cytosolic concentration of ions and the membrane
potential.

ligand-gated channels
 binding of an extracellular or
intracellular small molecule forces an allosteric transition
in the protein.

Voltage-gated ion channels


 a change in transmembrane
electrical potential (Vm) causes a charged protein domain
to move relative to the membrane.
 Ion channels provide hydrophilic pores through
which select ions can diffuse, moving down their
electrical or chemical concentration gradients; they
characteristically are unsaturable, have very high flux
rates, and are highly specific for one ion.

----------------------------END OF TRANS--------------------------

References:
Lecture’s ppt and recording

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