BIOC 2580 Chapter Notes - Chapter 4: Cyanogen Bromide, Reagent, Chymotrypsin

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Determining the amino acid sequence of a protein 1. Synopsis: in practice, proteins need to hydrolysed into shorter peptides for sequencing. Selective hydrolysis of the polypeptide chain by proteases, or with chemicals, cuts very long polypeptides into specific fragments of more manageable sizes. Using tandem mass spectrometry, proteins can be sequenced and their identity determined by searching protein databases and using search tools like blast. Selective hydrolysis of polypeptides allows a long polypeptide to be cut at specific locations, to give shorter oligopeptides. If the oligopeptides are no longer than 20-30 amino acids, their sequences can be determined by edman"s method or mass spectrometry. Selective hydrolysis can be achieved with the help of digestive enzymes called proteases. Enzymes are proteins that catalyze a specific reaction, in this case, hydrolysis of the targeted peptide bond. Trypsin is an enzyme that binds a polypeptide and cuts the peptide bond on the carboxylate side of the targets.

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