CRM proteins

Question 1

central dogma

Answer 1

genome(DNA to mRNA) to proteome (nascent polypeptide to mature protein to modified protein)
transcription and translation and post translational modification changes can alter protein state and function

Question 2

4 bases to 40 AA

diversity of production regulation

Answer 2

chromatin state of DNA region
mRNA stability and miRNA
translational efficiency
protein and mRNA have different decay rates

Question 3

mRNA and proteins

Answer 3

miRNA effect genes by targeting mRNA degradation and translation repression
ribosomal occupancy related to protein abundance

Question 4

Bicinchoninic acid assay (BCA assay)

Answer 4

determines protein conc in solution by colour change, used absorption’s vs known conc.

Question 5

single protein analysis
hybridoma cell lines
ELISA (enzyme linked immunosorbent assay)
PAGE gels

Answer 5

hybridoma cell lines gives permanent monoclonal antibody source
detects and qualifies antigen in sample, antibody binds, another antibody linked to enzyme binds and also binds to substrate initiating colour change
proteins+SDS gives size dependent negative charge then separated

Question 6

genetically engineered tags

Answer 6

epitope tagging for localisation and purification

GST tagged fusion protein purification using affinity chromatography

Question 7

study on UMOD expression

Answer 7

reduced UMOD reduces na absorption in TAL reduces ECV and BP

Question 8

gene variations and protein variations and drugs

Answer 8

rarely protein mutations effect protein folding but can effect PPI and drug protein interactions

Question 9

SNP and drug treatments
CYP2C9 and warfarin
CYP2D6 and codeine
herceptin and cancer

Answer 9

metabolises warfarin (anticoagulant) and 20% prescription drugs, too little warfarin ineffective, too much bleed risk eg brain, gene based prescription

some ethnic based codeine prescriptions due to differing levels of metabolisation

HER2 and cancers eg breast and stomach can attack molecular genetics of cancer

Question 10

proteome wide analysis (mostly too large)

Answer 10

proteomics is comprehensive quantitative description of protein expression and its changes under biological perturbations

Question 11

HPLC (high protein liquid chromatography)
normal chromatography
mass spec

Answer 11

AA seperation

ion or size separation

measures molecular mass can detect minor mass changes eg AA sub or pot T mod also immunoprecipitation can see which bit binded