Block B Workshop: Kinases

Question 1

What is reversible covalent modification?

Answer 1

Regulation of enzyme activity via covalent changes.

E.g Phosphorylation, Acetylation, Methylation, Ubiquitination etc… or the reverse of any of these.
(Slide 2)

Question 2

What are Phosphatases?

Answer 2

Enzymes that remove phosphate groups from proteins
(Slide 3)

Question 3

Why does phosphorylation change the 3D conformation of the target protein?

Answer 3

Because of the high charge density of the protein-bound phosphoryl group (-2 at physiological pH), with these often making salt bridges with nearby Arginine or Lysine amino acid residues which are positively charged
(Slide 4)

Question 4

What does protein kinase A phosphylate?

Answer 4

Amino acid residues on its target proteins that are part of a specific peptide region
(Slide 11)

Question 5

What are antibodies raised against the phosphorylated form of the specific peptide region which contains amino acid residues PKA phosphorylates called?

Answer 5

Phospho-PKA substrate antibodies
(Slide 11)

Question 6

Are phospho-PKA substrate antibodies man-made or naturally made?

Answer 6

Man-made
(Slide 11)

Question 7

What are phospho-PKA substrate antibodies used for?

Answer 7

They can be used to identify and study phosphorylated target proteins
(Slide 11)

Question 8

What occurs in in vitro phosphorylation reactions?

Answer 8

An active kinase is added to a cell extract and the consequences for protein phosphorylation can be studied
(Slide 11)

Question 9

What is a cell extract?

Answer 9

A solution containing cellular components for experiments
(Slide 11)

Question 10

What are in vitro phosphorylation reactions used for?

Answer 10

To identify kinase targets
(Slide 11)

Question 11

What is SDS-Page?

Answer 11

A technique for separating proteins by size.
(Slide 12)

Question 12

What is Western Blot Analysis?

Answer 12

A method to detect specific proteins using antibodies.
(Slide 12)