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BIOL 331 > Comprehension - L05-L06 > Flashcards

Comprehension - L05-L06 Flashcards

1
Q

What is a facilitative transporter?

A

Binding of the solute triggers a conformational change in the transmembrane protein that exposes the solute to the other side

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2
Q

What is an example of a facilitative transporter?

A

GLUT4
Continued diffusion of glucose in the cell is possible because it’s phosphorylated

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3
Q

What are 5 examples of passive transport mechanisms?

A
  1. Diffusion through membrane
  2. Voltage gated ion channel
  3. Facilitative transporter
  4. Osmosis
  5. Ligand gated channel
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4
Q

What type of energy is used in active transport?

A

Hydrolysis of ATP (Primary active transport)
Flow of other substances down their concentration gradients (secondary active transport

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5
Q

What are the 3 types of primary active transport?

A
  1. P-type pump
  2. V-type pump
  3. ABC transport
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6
Q

How does the P-type pump work?

A
  • Becomes phosphorylated during active transport
    -Contributes to maintaining the membrane potential voltage in cells
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7
Q

What is pumped in/out per one ATP in the Na/K-ATPase P-type ion pump?

A

3 Na+ pumped out and 2 K+ pumped in

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8
Q

What are the steps of the Na+/K+ -ATPase P-Type ion pump?

A
  1. High affinity for Na+ and ATP is bound
  2. Protein closes once ions are bound
  3. Hydrolysis of ATP and pump is phosphorylated
  4. Release of ADP –> E2. Loses affinity for Na+, high affinity for K+
    5-7. Dephosphorylation
    7-8. ATP binds,–>E1. Low affinity for K+
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9
Q

What are the characteristics of V-type ion pumps?

A

Use ATP but don’t become phosphorylated
Transport H ions across organelles and vacuoles
Also found in PM of some cells

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10
Q

What are the characteristics of an ABC transporter?

A

-ATP Binding Cassette transporters
- Share similar structure of ATP binding domain
- Transport ions, lipids, peptides, and nucleosides

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11
Q

In secondary active transport, what is a symporter?

A
  • Transports two substances in the same direction
  • Also called cotransporter
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12
Q

In secondary active transport, what is an anti porter?

A
  • Transports two substances in opposite directions
  • Also called an exchanger
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13
Q

What does the Na+/glucose cotransporter do?

A
  • Transport glucose from the lumen into epithelial cells
  • Na+ ions [ ] is low inside cells
  • Na+ ions moving down their [ ] gradient is used to drive the cotransport of glucose
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14
Q

What form of transport establishes the Na+ [ ] gradient in the first place?

A

Primary active P- Type

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15
Q

What is FRAP?

A

Fluorescence recovery after photobleaching
- Technique to study movement of membrane components

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16
Q

What are the 3 steps of FRAP?

A
  1. Label membrane w fluorescent dye
  2. Photobleach from a portion of cell
  3. Monitor reappearance of fluorescence in the previously bleached portion.
17
Q

What does the rate of recovery of fluorescence measure in FRAP?

A

Rate of diffusion of the fluorescently labeled portion

18
Q

When isolating membrane proteins, what is the first step? What happens in this step?

A

Lyse the cells!
Mechanical disruption (Hypotonic)
Pellet 1: insoluble membranes and proteins
Supernatant 1: soluble material

19
Q

When isolating membrane proteins what is the second step? What happens in this step?

A

Isolate peripheral proteins!
High salt solution
Pellet 2: transmembrane proteins
Supernatant 2: Peripheral proteins

20
Q

When isolating membrane proteins, what is the third step? What happens in this step?

A

Isolate Transmembrane proteins!
Strong detergent to break VDW interactions
Pellet 3: GPI-anchored proteins
Supernatant 3: Transmembrane protein

21
Q

Where are GPI-anchored proteins usually found?

A

In detergent resistant portions of the membranes that are rich in cholesterol and sphingolipids

22
Q

When isolating membrane proteins, what is the fourth step? What happens in this step?

A

Isolate GPI-anchored protein
Treatment w PI-PLC
Pellet 4: insoluble materials
Supernatant 4: GPI-anchored proteins

23
Q

When performing SDS-Page, how are proteins separated by size?

A

Adding SDS to the samples. This is a negatively charged amphipathic detergent that gives proteins a uniformly negative charge and denatures the protein

24
Q

What is the name of the dye that stains the proteins in polyacrylamide gel electrophoresis?

A

coomassie blue

BIOL 331 (10 decks)

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