Biochem

Front 1

biochemical assay

Back 1

measuring amount of biological substance in a complex mixture of other biomolecules

Front 2

amino acid residue

Back 2

-NH-C(R)H-C(=O)

Front 3

polypeptide

Back 3

-H3N+-CH(R)-C(=O)-NH-CH(R)-C=(O)-NH-CH(R)-C(=O)-

Front 4

nonpolar aa's

Back 4

glycine, alanine, valine, leucine, isoleucine, methionine, proline
(gladly allow valet lunatics into my protoge)

Front 5

aromatic aa's

Back 5

phenylalanine, tyrosine, tryptophan
(flaming t-rexes try)

Front 6

polar aa's

Back 6

serine, threonine, asparagine, glutamine, cysteine
(searing their asparagus;; glorious cynical)

Front 7

positively charged aa's

Back 7

lysine, arginine, histidine
(liars are historically)

Front 8

negatively charged aa's

Back 8

aspartate, glutamate
(assholes, mate)

Front 9

isoleucine

Back 9

ile, I

Front 10

phenylalanine

Back 10

phe, F

Front 11

tyrosine

Back 11

tyr, Y

Front 12

tryptophan

Back 12

trp, W

Front 13

asparagine

Back 13

asn, N

Front 14

glutamine

Back 14

gln, Q

Front 15

lysine

Back 15

lys, K

Front 16

arginine

Back 16

arg, R

Front 17

aspartate

Back 17

asp, D

Front 18

glutamate

Back 18

glu, E

Front 19

only non chiral aa

Back 19

glycine

Front 20

only imino acid

Back 20

proline

Front 21

phenylalanine wavelength

Back 21

258

Front 22

tyrosine wavelength

Back 22

275

Front 23

tryptophan wavelength

Back 23

278

Front 24

cysteine pka

Back 24

8.3

Front 25

lysine pka

Back 25

10.5

Front 26

arginine pka

Back 26

12.5

Front 27

aspartate pka

Back 27

3.7

Front 28

glutamate pka

Back 28

4.2

Front 29

histidine pka

Back 29

6

Front 30

SH-containing aa's

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methionine, cysteine

Front 31

NH-containing aa's

Back 31

tryptophan, lysine, arginine, histidine, asparagine, glutamine

Front 32

spectrophotomeric assay

Back 32

lactate dehydrogenase: Lactate -> pyruvate (C-OH -> C=O)

Front 33

immunochemical assay

Back 33

HIV coat proteins in human serum

Front 34

specific activity

Back 34

ratio of amount of desired protein present to amount total protein present in sample; should rise in purification

Front 35

lowry method for protein determination

Back 35

peptide bonds + copper in OH- + folin reagent -> blue

Front 36

bca method for protein determination

Back 36

protein + Cu(2+) > OH- > Cu(1+)

Front 37

bradford method for protein determination

Back 37

protein + coomassie -> blue

Front 38

UV absorbance for protein determination

Back 38

A = log Io/I = Ecl

Front 39

typical buffer components (6)

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buffering species
salt
reducing agent
chelator
protease inhibitor
other

Front 40

differential centrifugation

Back 40

separates different organelles of cells by repeated centrifuging at higher speeds

Front 41

dialysis

Back 41

removal of salt

Front 42

salting out effect

Back 42

using (NH4)2 SO4; aggregates broken apart proteins

Front 43

ion exchange chromatography

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in negative ions, most negative travel furthest

Front 44

affinity chromatography

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purification of recombinant proteins, specific ligand binding

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size exclusion chromatography

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stokes radius, balls with paths in it, largest come out first

Front 46

PAGE

Back 46

smallest molecules migrate furthest after electrophoresis; in bromphenol blue

Front 47

aa sequence determination

Back 47

- learn magnitude of problem
- detection of free aa's
- determine what is at end (amino)
- determine what is at carboxyl-terminal
- reduce disulfide bonds
- cleave with trypsin
- edman degradation
- peptide sequence determination by tandem MS
- establish sequence

Front 48

how to renature ribonuclease

Back 48

slow dialysis

Front 49

weak forces in 3D structure of protein (4)

Back 49

- hydrogen bonds
- electrostatic interactions
- van der waals attraction
- hydrophobic effect