BCHS 3304 Lecture Notes - Lecture 6: Cysteic Acid, Sanger Sequencing, Disulfide
21 Sep 2016
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Sanger sequencing can be used to find the order of aa in a protein: protein sequence is important because: Knowing the sequence will allow you to determine the structure. Comparing sequences can help figure out function and evolutionary relations. Mutations can lead to diseases, sequencing can help scientists find drugs to fix such mutations: steps to sanger sequencing. Fragment the pp into small peptides polypeptide chains: can use trypsin, chymotrypsin, chemical, cleave disulfide bonds separate polypeptides, prevents the protein from refolding, via performic acid oxidation. Cystine to cysteic acid: can use mercaptans that reduce cystine to cysteine, iodoacetate blocks the disulfide bond formation altogether. Determine aa composition of each peptide: determined by hydrolyzing the chain. This separates the aa: then you analyze the liberated amino acids, aa composition is indicative of protein structure. Ex: the ratio of polar to nonpolar tells you if its more likely a membrane or globular protein.
