Exam 1

Question 1

Name 4 main cell isolation methods

Answer 1

Tissue laser microdisection, immunoprecipitaion, differential centerfugation, flow cytometry with FACS

Question 2

FACS stands for

Answer 2

Florescence Activated Cell Sorting

Question 3

Flow Cytometry and FACS steps

Answer 3

cells are florescently labeled, suspended in individual droplets, ran through a laser and identified
+FACS, electrical charge is applied to the cells based on type, then used to sort into the appropriate tube

Question 4

Mass spec uses

Answer 4

used to identify protein expression levels and modifications

Question 5

Mass Spec types

Answer 5

1. Standard

2. Tandem

Question 6

Mass Spec process

Answer 6

ion charge is applied in a vacuum, protein is fragmented, fragments are displayed at each mass

Question 7

Protein analysis by ChIP is used

Answer 7

Used to detect protein bound to regulatory sequences

Question 8

ChIP steps

Answer 8

1. protein fixed to DNA with formaldehyde
2. cell is lysed
3. DNA is fragmented with sound waves or enzymes
4. precipitation using antibodies bind target proteins, and thus also the bound DNA
5. amplify precipitated DNA via PCR

Question 9

SH2

Answer 9

domain that binds phos Tyrosine residues

Question 10

SH2 domain is important for

Answer 10

cell signaling

found on adaptor proteins, signal transduction

Question 11

Src kinase has – domains

Answer 11

4

Question 12

Src kinase 4 domains do what?

Answer 12

2 - kinase activity
1 sh2 - recognize phospho tyrosine
1 sh3 - bind to proline rich peptides for signal transduction

Question 13

domain evolution driven by -

Answer 13

domain shuffling and gene duplication

Question 14

greater number of non covalent interactions — how well ligand will bind in binding site

Answer 14

increases

Question 15

Km is a measure of

Answer 15

conc at 1/2 Vmax, shows enzyme substrate affinity, lower = higher affinity

Question 16

2 ways phosphate groups can be used to regulate protein function

Answer 16

1. addition of P to OH group directly, reversible, can turn a protein on or off
   ex. activation of Src kinase
2. GTP binding proteins, P is attached or not to GTP, instead of directly to the protein

Question 17

Src kinase activation

Answer 17

1. removal of P to loosen conf,
2. activating ligand binds to SH3 domain
3. kinase phos tyrosine to self activate

Question 18

in lipid membrane, higher cis bonds =

Answer 18

greater fluidity

Question 19

3 types of transport that require no energy

Answer 19

1. passive diffusion
2. facilitated diffusion
3. filtration

Question 20

2 types of transport that require energy

Answer 20

1. pinocytosis

2. active transport

Question 21

types of drug transport depend upon

Answer 21

size, plolarity and hydrophobicity of the drug

Question 22

water soluble drugs can pass via

Answer 22

passive diffusion through aqueous channels

Question 23

lipid soluble drugs can travel via

Answer 23

hydrophobic binding with the lipid membrane = passive diffusion

Question 24

non soluble and non lipid drugs must pass through membrane using

Answer 24

active transport OR facilitated diffusion

Question 25

most drugs are —

Answer 25

electrolytes (weak/acids or bases)

Question 26

orally administered drugs must pass though

Answer 26

1. epithelium - must cross in and back out of these cells

2. endothelium - loosely packed lining of capillaries, can diffuse in between the cells passively

Question 27

Blood brain barrier is a result of

Answer 27

tightly packed endothelium of capillaries, prevents passive diffusion

Question 28

— of drugs cannot cross into brain

Answer 28

98%

Question 29

3 ways to circumvent blood brain barrier

Answer 29

1. highly lipophilic drugs can diffuse through
2. specialized transporters can be used
3. drug may be injected directly into the CSF

Question 30

way to get molecule into the brain?

Answer 30

make it lipophilic

Question 31

Mutant CFTR allele can cause (2)

Answer 31

1. less of the protein to be produces OR 2. a less effective variant of the protein to be produced

Question 32

CFTR defect CLASS I

Answer 32

no synthesis of the protein occurs

Question 33

CFTR defect CLASS II

Answer 33

protein processing issue, not folded correctly

Question 34

CFTR defect CLASS III

Answer 34

ion gating defect reduced channel opening

Question 35

CFTR defect CLASS IV

Answer 35

reduced channel conductance, so ion can’t pass through as easily

Question 36

CFTR defect CLASS V

Answer 36

splicing defect results in lowered synthesis

Question 37

CFTR defect CLASS VI

Answer 37

protein instability leads to higher turnover rate on the membrane

Question 38

CFTR defects classes one, two, five and six result in

Answer 38

reduced quality of the transporter

Question 39

CRTR defects three and four result in

Answer 39

reduced function of CFTR at the surface