TD: Proteomics 1

Question 1

What is proteomics?

Answer 1

The study of the proteome cia the systemic and global analysis of all proteins encoded within the genome.

Involves looking at the function and structure of proteins

Question 2

What is the central dogma?

Answer 2

Question 3

What differences occur to the mRNA after transcription in prokaryotic and eukaryotic cells?

Answer 3

Question 4

Describe more in depth about the RNA processing that occurs to eukaroytic primary mRNA

Answer 4

Question 5

What are the different structure of protiens?

Answer 5

Question 6

What are examples of tertiary and quaternary structures?

Why is the folding of proteins important?

What other processes can affect protein structure?

Answer 6

Tertiary - myoglobon

Quartenary - haemoglobin

Proteins must fold into their proper 3D structure before they are active

Post-translational events affect the structure, activity and destiation of proteins

Question 7

Glycosylation is a major post-translational modification. Describe it

Answer 7

Question 8

Functions of glycosylation?

Answer 8

Question 9

Describe phosphylation

How does it occur

What is its function?

How are Pi added?

Answer 9

Protein kinases add Pi and phosphatases remove Pi. Adding or removing can either switch on or off a Pw

Tyrosine, serine and theronine are phosphrylated AA

Question 10

What are the 2 important regions of an AB?

What are each of their functions?

Answer 10

Epitope - part of antigen of foregin molecule with AB binds too

Question 11

Describe where AB glycosylation occurs and why it is important

Answer 11

Question 12

One gene —> One protein

Our DNA encodes about 20,000 different genes. Where do the millions of differen antigen receptor proteins come from? One gene generates many receptors? How?

What are the mechanisms for generating AB diveristy?

Answer 12

Summary of mechanism for generating AB diversity:

• Somatic recombination of light chain: V to J recombination
• Somatic recombination of heavy chain: D to J, and then V to DJ recombination
• Multiple V, D and J segments
• Junctional diversity

1. Light Chain Gene contains multiple V and Multiple J segments:

• Light chains can be lamda or kappa - the chain will only ever contain only kappa or only lamda
• The light chain has multiple J - junction regions (1-5) and V - Variable regions (up to 40)
• Any combination of one V and one J segment results in one light chain

2. Multiple segments of V,D and J of heavy chain gene

• The heavy chain contains multiple V - Variable (up to 65 types), D - Diversity (up to 27 types) and J - Junction (up to 6 types).
• Any combination one one V, one D and one V results in one heavy chain
• 5 different types of heavy chain:
  
  • alpha - IgA
  • Epsilon - IgE
  • Mu - IgM
  • G - IgG
  • D - IgD

3. Junctional Diveristy

Can get even more diveristy when segments are joined together. Depending on where the segments join e.g. V and J in the light chain determines the codon produced therefore the AA encoded for.

See next few slides for pictures of each

Question 13

Light chain diveristy

Answer 13

Heavy chain diversity

Question 14

Junctional diveristy

Question 15

Describe the binding of AB to antigens:

Consider:

• Where bind to?
• Multiple binding?
• How bind?

Answer 15

Question 16

Describe the different and use of polyclonal AB and monoclonal AB

Answer 16

Question 17

Titers and dilutions

Answer 17

Question 18

Applications of mAB

Answer 18