lecture 7- proteins

Question 1

what is the main purpose of solid phase peptide synthesis?

Answer 1

to build a polypeptide chain

Question 2

in spps, what is used to activate the carboxyl groups for the formation of peptide bonds?

Answer 2

dicyclohexylcarboiimide (DCC)

Question 3

what does spps start with?

Answer 3

immobilizing the c terminal amino acid of the target peptide sequence on a resin

Question 4

second t-boc amino acid is added and coupled using what chemistry?

Answer 4

carbodiimide

Question 5

carbodiimide does what?

Answer 5

reacts with and activates the carboxyl group of the t-boc AA to facilitate peptide bond formation

Question 6

during spps, what blocks the amino ened?

Answer 6

t-boc

Question 7

first amino acid will add to what end on the spps process?

Answer 7

carboxyl

Question 8

t boc is removed by what in spps?

Answer 8

trfluoroacetic acid

Question 9

what carries amino acid in spps?

Answer 9

polystyrene bead

Question 10

when done with spps, what do you od to get rid of bead?

Answer 10

chemically cleaves

Question 11

explain indirect ELISA

Answer 11

you first coat the wells with antigen, then add specific antibody , then enzyme linked antibody binds to specific antibody. substrate added and converted by enzyme into colored product.

Question 12

in indirect ELISA, rate of colo formation is proportional to amount of what?

Answer 12

specific antobody

Question 13

explain sandwhich ELISA

Answer 13

MONOCLONAL ANTOBODY-COATED WELL, ANTIGEN BINDS TO ANTIBODY, SECOND MONOCLONAL ANTIBODY, LINKED TO ENZYME, BINDS TO IMOBILIZED ANTIGEN

Question 14

in sandwich elisa, rate of color formation is propotional to amount of what

Answer 14

anitgen

Question 15

elisa is mostly responsable for doing what?

Answer 15

quantities of proteins

Question 16

which elisa process is more specific?

Answer 16

sandwich

Question 17

name some of the advantages of elisa

Answer 17

extremly sensitive to detection of antibody and antigen. able to have multiple wells, quantitative readout of antigens or antobody level in a fluid. there are many tag s available. many commerical kits available

Question 18

disadvantes of elisa?

Answer 18

would need to have specific antibodies, conditions are specific to the pair of antibody and antigen. very time consuming, and expensive.

Question 19

explain what western blotting is mostly used for

Answer 19

ability to find out if certain protein is present

Question 20

explain problem with western blotting

Answer 20

not all antibodies have high enough affinity

Question 21

in order to use western blotting, what do you have to do?

Answer 21

cut out of sds gel

Question 22

how many human genes?

Answer 22

30k

Question 23

what is functional genomics? focuses on what?

Answer 23

affort to make use of the vast wealth od data from the various genomics projects to understand gene and protein functions and interactions. focuses on transcription, translation, and protein-protein interaction

Question 24

glycomics

Answer 24

study of glycosylation of proteins and lipids on an organismal scale

Question 25

metabolics

Answer 25

study of the patterns of small molecules metabolites expressed in a biological system

Question 26

circular dichroism is useful for what?

Answer 26

useful for finding if protein has been modified

Question 27

light in circular dichorism is what?

Answer 27

circular polarized light

Question 28

schemes of the electric field components of unpolarized give what on the graph

Answer 28

multiple lines

Question 29

plan polarized light gives what?

Answer 29

one line

Question 30

circular dichorism allows you to differ from what two things?

Answer 30

beta and alpha

Question 31

explain to different graphs from CD

Answer 31

near Uv, and far Uv.

Question 32

near UV and far UV from cd allow you to determine what?

Answer 32

comparing secondary structure, normal version vs mutant, use as a way of tudying kinetics. native proteins will absorb differently

Question 33

how do you determine melting point of proteins?

Answer 33

melt down into primary structure and then test through CD

Question 34

Nmr allows you to do what?

Answer 34

helps determine functional groups and create 3d structure. determines distances between pairs of atoms.

Question 35

in NMR, what determines the environment of protond and other nuclei?

Answer 35

nuclear magnetic resonance

Question 36

NMR is limited to what?

Answer 36

small proteins, cannot do very large proteins

Question 37

NMR is a form of what?

Answer 37

spectroscopy