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Review Sheet 3 - Katja Dove Biochem 440 Review Sheet...

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Katja Dove Biochem 440 Review Sheet 3 [GdnHCl] 50% = concentration of GdnHCl at which 50% protein unfolded T m = Temperature at which 50% of protein is unfolded *Molecular chaperons* : protein helpers proteins that interact with partially folded or improperly folded polypeptides Heat Shock proteins(rescue) bind solvent-exposed hydrophobic regions of unfolded P (prevent aggregation) Chaperonins ( helpers ; require more Energy than heat shock proteins ) Help proteins that don t fold spontaneously on their own *Protein Purification + Visualization * Isoelectric precipitation/Salting out o using pI (Net charge of protein is zero) minimal solubility in solution precipitate, separate from soluble proteins by centrifugation o pI-pH<0 protein negatively charged o pI-pH>0 protein positively charged Column Chromatography o Separate mixtures of proteins according to properties Size Size Exclusion Chromatography (Gel filtration) Charge Ion Exchange Chromatography
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