BISC 330 Spring 2010 Lecture 9

BISC 330 Spring 2010 Lecture 9 - BISC330L:Biochemistry...

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BISC 330L:  Biochemistry Spring Semester 2010 USC Lecture:  Monday, 01 February 2010 1. Chapter 3:  Biochemistry Toolbox Techniques in Protein Analysis Determining Protein Sequence
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SDS-PAGE Revisited • The most powerful technique for  resolving protein mixtures. • Proteins are exposed to the ionic  detergent SDS (sodium  dodecylsulfate)  before and during  electrophoresis.  SDS is an ionic detergent that is able  to denature proteins (disrupts weak  non-covalent interactions)   β -mecaptoethanol  is added to  protein samples…to break/reduce  disulfide bonds
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SDS-PAGE: Denaturing Gel Extended conformations with similar  charge:mass ratios • SDS denatures proteins, causing  multimeric proteins to dissociate  into their individual subunits • SDS coats the all polypeptide  chains with a  negative charge ,   therefore forcing the polypeptide  chains into extended conformation  with  similar charge:mass ratios • SDS eliminates the effect of  differences in shape, and so  chain   length (mass) is the sole  determinant of the migration  rate in SDS-gel electrophoresis
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sequences important?   Can make comparisons to other known  proteins – Proteomics Reveal evolutionary pathways Search for internal repeats Can say something about destination or 
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This note was uploaded on 08/30/2010 for the course BIO 320 taught by Professor Wu during the Spring '10 term at USC.

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BISC 330 Spring 2010 Lecture 9 - BISC330L:Biochemistry...

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