Lecture 10: Protein Analysis

Question 1

the best analysis of enzymatic function is done with __- protein

Answer 1

pure

Question 2

what are the 3 general steps to protein purification?

Answer 2

1. disrupt the cells to release proteins
2. separation techniques that exploit differential properties
3. monitor success with gel electrophoresis and staining techniques

Question 3

cell lysis involves breaking open cells while leaving __ and ___ intact

Answer 3

cell membrane; organelles

Question 4

a cell extract obtained after lysis is called ___

Answer 4

homogenate

Question 5

extraction of the protein pool can be done by differential ___

Answer 5

centrifugation

Question 6

what are the 3 main column chromatography techniques?

Answer 6

1. ion exchange chromatography
2. affinity chromatography
3. size exclusion chromatography

Question 7

ion exchange separates proteins on the basis of their __

Answer 7

charge

Question 8

affinity chromatography separates proteins on the basis of the ability of certain proteins to ___

Answer 8

bind to certain other molecules (ligands)

Question 9

size exclusion chromatography separates proteins on the basis of ___

Answer 9

size

Question 10

in column chromatography, a mixture of proteins is carried in a ___ solution through a __

Answer 10

buffer; porous column of small beads

Question 11

in column chromatography , the mix of proteins is separated by ____ interactions with the matrix

Answer 11

differential

Question 12

the more a protein interacts with the matrix, the __ it will elute from the column

Answer 12

later

Question 13

proteins with a pI above the pH has a net __ charge

Answer 13

+

Question 14

proteins with a pI below the pH has a net ___ charge

Answer 14

-

Question 15

positively charged proteins are called __ proteins

Answer 15

basic

Question 16

negatively charged proteins are called __ proteins

Answer 16

acidic