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21 Cards in this Set
- Front
- Back
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proteome***
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name given to the full complement of proteins or cell or organisms produce
-proteomics-study of this field DNA level (genomics) Rna level (transcriptomics) proteins level (Proteomics) metabolites level (metabolomics) |
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why proteomics
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genome analysis look at DNA seq
transcriptome analySis look at expression data ???these analysis x same as looking at actual protein data -gene seq -mature RNA -functional protein |
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???Proteomics
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proteins exp level x predictable from mrna exp level
uniquely modified n processed different than gene seq Proteoms are dynamic |
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phosphorylation
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signal transduction
gene expression cell division |
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ubiquitination
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ubiquitin - 8.5 kDa protein in all eukaryotic cells tagging old, damaged or,defective protein for degradation
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glycoprotein cell receptor
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surface carbohydrate as point of attachment
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glycosylation
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addition of short chain carbohydrates residues to protein during@after,synthesis
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definition proteomics
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branch of biotech
protein produced by gene info in database application of data |
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Qs
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??analyze protein
??proteome can we predict proteome of cell from genome of transcriptome |
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differential precipitation
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precipitation - achieves separation by conversion of soluble protein to an insoluble state, which can be subsequently be removed by various means
when ammonium sulphate @ polyethylene glycol are added to a protein, a precipitate forms & it can be,separated from solution after centrifuge |
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gel filtration chromatography
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briefly separate protein according to size
beads,got pore small sizes trapped inside,beads get samples with large,sizes protein x accurate |
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gel filtration chromatography
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gel permeation/size exclusion
separate acc molecular,size adv-large quantity protein can be separated |
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ion exchange
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each protein separated based on.charge protein
DEAE cellulose -'ve charge carboxymethyl cellulose - +ve charge |
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reversed phase hplc
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octacecyl
butyl propyl phenyldimethyl -many column sizes with diff diameter |
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affinity chromatography a)
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enzyme - substrate
antibody - antigen receptor - ligand #knowledge about protein essential exp - concavAlin A is glucose binding protein ~ can be purified by passung thorough column that has beads which have glucose attached to them ~protein binds specifically to ligand attached to matrix |
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affinity chromatography
b) |
antibody affinity chromatography
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What is structural proteomics?
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high throughput determination of 3d structure protein
goal- |
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xray crystallography
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make crystals ur protein
protein must crystallize need to have access to x Ray beam solving structure |
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predict structure polypeptide by bioinformatic
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comparative modelling BLAST
threading/fold recognition Ab initio |
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classification scheme
structural proteomics direct method determination protein |
note****
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iTRAQ
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isobaric tags for relative and absolute quantitation
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