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1/31/20171ProteinsPart 3CharacterizationProtein CharacterizationThe protein has been purified, what’s next?Questions:•How is the methane produced?•What amino acids are important for this activity?•What does the active site looks like?•Is there a cofactor present/other modifications?Reaction mechanismOptions:•Study reaction kinetics (enzymology notes)•Get amino acid sequence•Determine exact mass of protein•Compare theoretical mass (from sequence) with actual mass•Determine 3D structure (Part 1 Notes)33.1)Sequencing Procedure1) Obtain amino acid composition2) Determine which amino acid is present at the N-terminus3) Break disulfide bonds if present4) Sequencing4
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