BISC 330 Spring 2012 Lecture 9

BISC 330 Spring 2012 Lecture 9 - BISC 330L Biochemistry...

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BISC 330L: Biochemistry Spring Semester 2012 USC Lecture 9: Monday, 30 January 2012 1. Chapter 3: Biochemistry Toolbox Techniques in Protein Analysis Determining Protein Sequence 3 0 L
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Why are amino acid sequences important? Can make comparisons to other known proteins – Proteomics Reveal evolutionary pathways Search for internal repeats Can say something about destination or processing of protein Prepare antibodies – allow for further analysis Can make DNA probes
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Determining amino acid composition Peptide hydrolyzed in 6M HCl at 110 o C for 24 h Ion exchange chromatography Identity revealed by elution volume Quantity revealed by Ninhydrin (10 nmol) Can also use Fluorescamine (10 pmol)
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Protein Sequence Determination: Edman Degradation It is useful to know the specific amino acid sequence of a protein Edman Degradation: method developed in the 1950s by Pehr Edman to determine protein sequence With genome sequencing, this method is not as widely used. Protein sequence can be deduced directly from the DNA sequence based on The Genetic Code
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Edman Degradation
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BISC 330 Spring 2012 Lecture 9 - BISC 330L Biochemistry...

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