CH 13 Membrane Channels and Pumps P1 Flashcards
Membranes are made permeable to specific molecules in the presence of three classes of transporters:
ATP-driven pumps
carriers
channels
Lipophilic / Hydrophobic Molecules
pass directly through phospholipid bilayers down [gradient]
Polar / Fully Charged Molecules
req. channel to move across membrane down [gradient]
Simple diffusion
doesn’t req. E or membrane proteins
[high] to [low]
Facilitated Diffusion / Passive Transport
polar / fully charged molecules that req. presence of channel to move across a cell down [gradient]
Active Transport
moves molecules across membrane against [gradient].
req’s E
E-rich Situation
unequal distribution of uncharged molecule across membrane
Gibb’s Free E Eqn
dG = RT ln (c2/c1) R: gas constant T: temp K c1: [side] 1 c2: [side] 2
Gibb’s Free E
Free E req’d to move solute from one side of a membrane at [c1] to the other at [c2]
Un= distribution of a charged molecule has?
electrical potential
Electrochemical / Membrane Potential
concentration diff’s and charge diff’s taken into account for un= distribution of charged molecule w/ electrical potential
Electrochemical / Membrane Potential Eqn
dG = RT ln (c1/c2) +ZFdV
Z: charge of molecule
dV: voltage across membrane
F: Faraday constant
Most animal cells have ____ K+ and ____ Na+ relative to the environment.
high [K+]
low [Na+]
Na+K+ Pump
req’s action of active transport system to gen and maintain membrane potential
Na+K+ ATPase
P-type ATPases
phosphorylated aspartate intermediate
Na+K+ Pump
P-Types ATPases Couple ____ and ____ to pump ____ across membranes.
phosphorylation
conformational changes
calcium ions
Pumps exist in what 2 conformations?
ion binding site facing into cell and facing out of cell
ATP Hydrolysis
powers interconversion of 2 conformation
Sarcoplasmic Reticulum Ca2+ ATPase (SERCA)
P-Type ATPase
pumps Ca2+ from muscle cytoplasm to sarcoplasmic reticulum.
enzyme.
calcium pumping.
SERCA Structure
single polypeptide chain
transmembrane domain - 10 alpha helices
SERCA Function
binds Ca2+
SERCA Cytoplasmic Portion
3 Domains:
N domain
P domain
A domain
SERCA N domain
binds ATP
SERCA P domain
accepts P during rxn cycle
Asp 351 binds P
SERCA A domain
links N and P domains
Catalytic / Transport Cycle
- bind cytoplasmic Ca2+
- N domain binds ATP
- P transferred to Asp in P domain
- ADP released, structural change, Ca2+ binding site faces SR lumen, Ca2+ leaves enzyme
- P group in P domain hydrolyzed
- enzyme changes conf. Ca2+ binding site faces cytoplasm
Digitalis
inhibits Na+K+ pump by blocking dephosphorylation
Cardiotonic Steroids
found in foxglove.
inhibits Na+K+ ATPase
treats congestive heart failure.
Cardiotonic Steroids: Digitoxigenin and Ouabain
members of class of inhibitors
Digitalis decreases the ____ which results in ____.
Na+ gradient.
slower Ca2+ removal from cell.
Digitalis increases the ____ which results in ____.
cellular Ca2+.
enhances contractile ability of heart.
The human genome contains 70 genes that encode?
P-type ATPases
P-Type ATPases transport
H+ Ca2+ Na+ Cu2+ phospholipids w/ charged head groups
All P-Type ATPases have the same fundamental rxn mech:
take advantage of free E release associated w/ ATP hydrolysis to drive membrane transport via conformational changes induced by addition / removal of phosphoryl group at key Asp site.
Multidrug Resistance Protein
membrane pump. ATP-dependent pump. extrudes small molecules from cell. member of family - ABC transporters. single polypeptide chain w/ 4 domains.
ABC transporters are characterized by a common domain called?
ATP-binding cassette (ABC)
ABC transporters consist of 2 ____ and 2 ____.
ABC domains
membrane-spanning domains
Reaction Cycle of Multidrug Resistance
- channel open to cytoplasm
- substrate binds - conf. changes in ABC domain.
- ATP binds to ABC domains - structural changes orient substrate to face outside of cell.
- outward facing conf. to transporter reduced affinity to substrate - allows release.
- ATP hydrolysis - resets transporter to initial state.
Lactose Permease
from E.coli.
symporter.
uses H+ gradient to power lactose into cell.
Secondary Transporters / Cotransporters
use E of one gradient to power formation of another
Symporters
power transport of molecule against [gradient].
couples movement of another molecule down [gradient].
both molecules move in same direction.
Antiporters
use 1 [gradient] to power formation of another.
molecules move in opp directions.
Uniporters
transport molecule in either direction.
depend on [difference] across membrane
Lactose Permease Mech
- lactose-binding pocket faces outside of cell.
- H+ binds, lactose binds.
- permease everts.
- lactose leaves permease, enters cell.
- H+ leaves permease, enters cell.
- permease everts - cycle complete
Ion Channels
rapid movement of ions across membranes down [gradient]
Neuron Interior
higher [K+]
lower {Na+]
membrane potential -60mV
Nerve Impulse / Action Potential
gen’d when membrane potential depolarized beyond critical threshold value.
Action Potentials are the result of?
K+ channels and Na+ channels opening / closing in response to changes in membrane potential
Patch-Clamp Technique
pipette placed against small portion of cell membrane.
reveals activities of single channels.
Patch-Clamp uses Gigaseal which is?
suction applied to form tight seal btw pipette - plasma membrane.
measures channel activity when voltage applied across cell membrane.