Week 9 - Membrane Transport Pt II Flashcards
What does K ion channel coming of?
4 subunits
What does each subunit comprise of?
M1 and M2 helixes joined by a P (pore) segment consisting of a short helix and non helical portion that lines the channel through which ions pass
What does a portion of each P segment contain?
Conserved pentapeptide (GYGVT) whose residues kind the selectivity filter that screen for K+ ions
What does the selectivity filter contain?
4 rings of carbonyl O atoms and one ring of threonyl O atoms
Why is the diameter of ring large enough?
8 O atoms can coordinate a single K ion, replacing it’s normal water of hydration
Voltage-gated channels
Conformation depends on the difference in ionic charge on two sides of membrane
Ligand gated channels
Conformation depends on binding of a ligand to channel
ACH binds on outside and CAMP binds on inside
Mechano-gated channels
Conformation depends on mechanical forces applied to membrane
What are voltage-gated K+ channel from Eukaryotes known as?
Kv channels
What is the structure of KV channel?
Four identical/ homologous subunit
Each subunit: 6 transmembrane alpha helices
What are 2 functionally distinct domain of KV channel?
Pore domain
Voltage sensing domain
What is pore domain?
Helices S5, S6 and P segment homologous to helices M1, M2 and P segment in Kcsa
What is voltage sensing domain?
Consist of helices S1 to S4
How many subunits do all channels have?
4
Prokaryotic channel
4 subunits and 2 helices each=8
What is the role of transmembrane portion?
Voltage gating?
What is the role of cytoplasmic portion?
Regulation
What does amino acid sequence of S4 helix show?
A number of positively charged amino acid residues
What does S4 segment show?
A series of positively charged amino acids (Lysine and arginine) and every 2 residues, +ve side chain going down
Where does helix face when there’s an active membrane potential?
Cytoplasmic
More negative inside cell than outside
Why is S4 rotates?
Positive residues face electronegative area in cytosol
When does helix face outside of cell?
Losing negative charge on inside
What is transfixed to open channel?
Structural change
What does moving S4 helix transducer?
Movement of two helices (S4-S5) that forms channel for opening and closing of channel
What connects the voltage sensing and pore domain?
S4-S5 linker helix
What is the inner surface of channel below pore domain lined by?
S6 helix
Resting condition, negative potential
S4 helix exposed to cytoplasm
Channel closed
Depolarisation
S4 helix exposed to the outside of cell
Channel open
Once the channel is open, how many K+ ions can pass through per second?
10 million K+ ions
What is the conformational states of Kv channel?
Rest
Open
Inactivated
Conformational state: rest
Membrane potential is there
Pore is closed
Conformational state: open
Depolarising membrane
Flux of sodium
Conformational change: inactivated
Milliseconds after K+ channel shut down
Why do we need so many voltage gated K+ channel?
Depending on the charges on the S4 helix, it will rotate channel at higher potentials/lower potentials
Channel to detect subtle changes in membrane potential (differentiate to transduce lots of signals)
Switched on and off through: phosphorylation, PTM etc
What requires a facultative transporter to cross Membranes?
Large or hydrophilic substances
What is facilitative diffusion?
Passive Specific Saturable Regulated Much slower than ion channels
Faciliative transporters can move how much solute molecules per second across the membrane?
Hundred to thousands
What is a gradient favouring the continued diffusion of glucose into the cell maintained by?
Phosphorylating the sugar after it enters the cytoplasm
Lowers the intracellular glucose concentration
Transport
- Glucose binds to transporter
- Transporter changes shape
- After binding, active site now exposed to cytosolic side
- Transport
Recovery
Active site closed to cytosol
How many glucose transporters in human body?
5
Which glucose transporter is most important?
GLUT 4
Why is hexokinase phosphorylation?
Concentration gradient to drive glucose in
What happens when you eat a lot of sugar?
Pancreas produces insulin
Vesicles full of GLUT4 transporters on surface of cells
Vesicular fusion
Glucose import from bloodstream to cells (skeletal muscle and adipocytes)
Where are transporters present?
Membrane of cytoplasmic vesicles
What is nitrate central in?
Nitrogen metabolism
How is nitrite rapidly removed from the cell?
Channels
Transporters
What do assimilatory enzymes do?
Reduce nitrite to ammonium it dinitrogen
What is the structure of Nark?
Positively charged substrate-translocation pathway lacking protonable residues
What does Nark function as?
Nitrate/nitrite exchanger
Protons are unlikely to be co-transported
How does Nark exchange nitrate for nitrite by?
Rocker switch mechanism
Facilitated by: inter-domain hydrogen bond networks
Active Transport
Energetically unfavourable so needs to be coupled to a favourable reaction
What does active transport do?
Maintain gradients for: Potassium Sodium Calcium Other ions across cell membrane
Active transport
Couples the movement of substances against gradients to ATP hydrolysis
What is the ratio of Na+ : K+ pumped?
3:2
The Na+/K+ ATPase is electrogenic
What is ATPase?
P-type pump
Phosphorylation causes change in structure and affinity that allow transport against gradients
Where is Na+/K+ ATPase found only in?
Animals
What does hydrolysing ATP give?
Structural change in ATPase protein
Define electrogenic?
Producing a change in the electrical potential of a cell
What does Na+/K+ ATPase have high affinity for?
Na+ in the cytoplasm
K+ in the extracellular space
What does Na+/K+ ATPase have lower affinity for?
Na+ in the extracellular space
K+ in the cytoplasm
E1 —> E2
Phosphorylated and release ADP
E2 —> E1
Dephosphorylated and binds ATP
Where does sodium ions bind protein?
Inside of membrane
When ATP is hydrolysed andphosphate transferred to protein what does it allow?
Sodium ions to be expelled to external space
What happens to the phosphate group when potassium ions bind to the protein
Subsequently lost
Where is cation binding site located?
Deep within transmembrane domain
Consist of 10 membrane spanning helices
What does V-type pump actively transport?
Hydrogen ions across the walls of cytoplasmic organelles and vacuoles
Where does V type pump occur?
Membrane that line:
Lysosomes
Secretory granule
Plant cell vacoules
V type pump in the plasma membrane of kidney tubules
Maintain body’s acid-base balance by secreting protons into forming urine
What is active transport coupled to?
Existing ion gradient
What is symport?
Pump gradient up on inside
Move sodium with other compounds to create smaller gradients of other molecules
The molecules will travel in same direction in relation to each other
What is antiport?
Integral membrane transport protein that simultaneously transport two different molecules in opposite directions
Sodium glucose transporter
High sodium concentration outside
Low concentration of glucose
What is secondary transport?
The use of energy stored in an ionic gradient