Term
| Four ways to extract proteins from a cell |
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Definition
1. Sonication
2. French Press
3. Freeze/Thaw
4. Enzymatic |
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Term
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Definition
| Method for the estimation of protein concentration based on Cu2+ reduction |
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Term
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Definition
| A method for estimating protein concentration based on Coomassie Brilliant Blue G250 absorption |
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Term
| Ion Exchange Chromatography |
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Definition
| Ionic strength (salt) is used to separate proteins |
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Term
| Gel Filtration Chromatography |
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Definition
| larger particles travel more quickly through gel beads |
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Term
| Three factors affecting gel filtration chromatography |
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Definition
1. length of column
2. sample size
3. flow rate |
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Term
| SDS-PAGE Gel Electrophoresis |
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Definition
| electrophoretic separation of denatured proteins based on mass |
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Term
| Two-Dimensional Gel Electrophoresis |
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Definition
| Separation by IEF and then SDS-PAGE |
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Term
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Definition
| Use of substrates to separate proteins |
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Term
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Definition
enzyme activity (micromoles of S->P/min) per mg of enzyme
total activity/total protein |
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Term
| Six steps for determining protein sequence |
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Definition
1. Separate and purify chains
2. Cleave intrachain S-S bonds
3. Identify N-terminal and C-terminal residues
4. Cleave chains into smaller fragments and ID sequence
5. Repeat four with a different method
6. Reconstruct overall sequence |
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Term
| Methods for chain separation |
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Definition
| extreme pH, urea, guanadine HCl, high salt concentration |
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Term
| Methods for disulfide bridge cleavage |
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Definition
| performic acid oxidation, sulfhydryl reduction (BME) followed by iodoacetate |
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Term
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Definition
| used to ID N-terminal residues |
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Term
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Definition
| cleaves any residue except Arg, Pro, Lys |
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Term
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Definition
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Term
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Definition
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Term
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Definition
| cleavage after Phe, Tyr, Trp |
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Term
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Definition
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Term
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Definition
cleaves after Glu, Asp in phosphate buffer
Cleaves after Glu in acetate or bicarbonate buffer |
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Term
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Definition
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Term
| Electrospray Ionization (ESI-MS) |
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Definition
| Formation of highly charged droplets containing protein and consequent determination of m/z in MS |
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Term
| Mass, charge, and m/z relationship |
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Definition
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Term
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Definition
| a given protein is homologous across multiple organisms |
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Term
| Solid Phase Synthesis of Peptides |
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Definition
| Carboxy terminus is bound to a resin and residues are added in a stepwise manner |
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