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DMD biochem > lecture 7- proteins > Flashcards

lecture 7- proteins Flashcards

1
Q

what is the main purpose of solid phase peptide synthesis?

A

to build a polypeptide chain

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2
Q

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

A

dicyclohexylcarboiimide (DCC)

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3
Q

what does spps start with?

A

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

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4
Q

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

A

carbodiimide

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5
Q

carbodiimide does what?

A

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

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6
Q

during spps, what blocks the amino ened?

A

t-boc

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7
Q

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

A

carboxyl

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8
Q

t boc is removed by what in spps?

A

trfluoroacetic acid

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9
Q

what carries amino acid in spps?

A

polystyrene bead

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10
Q

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

A

chemically cleaves

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11
Q

explain indirect ELISA

A

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.

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12
Q

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

A

specific antobody

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13
Q

explain sandwhich ELISA

A

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

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14
Q

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

A

anitgen

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15
Q

elisa is mostly responsable for doing what?

A

quantities of proteins

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16
Q

which elisa process is more specific?

A

sandwich

17
Q

name some of the advantages of elisa

A

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

18
Q

disadvantes of elisa?

A

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

19
Q

explain what western blotting is mostly used for

A

ability to find out if certain protein is present

20
Q

explain problem with western blotting

A

not all antibodies have high enough affinity

21
Q

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

A

cut out of sds gel

22
Q

how many human genes?

A

30k

23
Q

what is functional genomics? focuses on what?

A

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

24
Q

glycomics

A

study of glycosylation of proteins and lipids on an organismal scale

25
Q

metabolics

A

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

26
Q

circular dichroism is useful for what?

A

useful for finding if protein has been modified

27
Q

light in circular dichorism is what?

A

circular polarized light

28
Q

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

A

multiple lines

29
Q

plan polarized light gives what?

A

one line

30
Q

circular dichorism allows you to differ from what two things?

A

beta and alpha

31
Q

explain to different graphs from CD

A

near Uv, and far Uv.

32
Q

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

A

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

33
Q

how do you determine melting point of proteins?

A

melt down into primary structure and then test through CD

34
Q

Nmr allows you to do what?

A

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

35
Q

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

A

nuclear magnetic resonance

36
Q

NMR is limited to what?

A

small proteins, cannot do very large proteins

37
Q

NMR is a form of what?

A

spectroscopy

DMD biochem (7 decks)

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