Lecture 23 - Proteomics Flashcards
How does SDS-PAGE analyze cellular proteins?
Separates on mass, smaller particles run further, does this by creating an equal charge across samples
How does IEF analyze cellular proteins?
Separates based on Charge!
How does Mass Spectrometry analyze proteins?
Ionized samples break into fragments that have a distinct mass to charge ratio. This ratio can be compared to known ratios to piece together molecule.
How does Western Blot work? (4 steps)
- Extract Proteins
- Electrophorese
- Transfer to membrane
- Detect with protein specific antibody
Protein microarrays are used for what?
Diagnostics, protein expression profiling, protein functional analysis, antibody characterization, and treatment development
What is Affinity capture?
A method of separating biochemical mixtures based on a highly specific interaction
Ie) antigen and antibody
Two-hybrid system is for what?
transcription factors. The transcription factors are split into two pieces but still work!!
What is an Interactome?
The whole set of molecular interactions in a particular cell.
What is Post translation modification?
Refers to the covalent and generally enzymatic modification of proteins following protein biosynthesis.
What are Prions?
infectious agents composed entirely of a protein material that can fold in abstract ways, that can be transmissible and lead to disease. Thought to cause spongiform encephalopathies.
What are genetic prion diseases caused by? what type of mutations are they typically?
Autosomal-dominant mutations.
Most PRNP mutations are missense (amino acid changes)
What are two protein databases used for structural/clinical prediction?
SIFT + PolyPhen-2
What are two protein functional databases?
Uniprot + Phyre
