Lecture 29

Lecture 29 - Lecture 29 Purification, detection and...

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Lecture 29 Purification, detection and characterization of proteins II Protein Sequencing Goal : You have a protein and you want to analyze its sequence Method : 1.Starting at the N-terminus you modify it 2.Cleave modified N-terminus 3.Analyze using high pressure liquid chromatography Concept : Each amino acid will migrate differently on a different column Problem : This type of sequencing loses its quality with time Only sequences for the first few amino acids are accurate One piece of the sequence is known and can be used to search in DNA sequence databases to determine the whole sequence Page | 1
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Mass Spectroscopy Principle: 1.Take a molecule and you break it with very high energy: 2.Covalent bonds are broken randomly to make charged fragments 3.Pieces of + charged molecules are attracted by a magnet to the target 4.A second magnet deviates the trajectory of these ions according to its weight Very precise: 1 atm is non ambiguously distinguished Tell difference between 37 and 38 atm Page | 2
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This note was uploaded on 06/05/2011 for the course BIO 200 taught by Professor Thomasebureau during the Fall '07 term at McGill.

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Lecture 29 - Lecture 29 Purification, detection and...

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