BIOS 452 Lecture Notes - Lecture 10: Hydrolysis, Random Coil, Carboxypeptidase

• School: UIC
• Department: Biological Sciences
• Course: BIOS 452
• Professor: Min Jung Hyun

Document Summary

Defined, non-random sequence of amino acids along the peptide backbone: described in two ways: Treat with 6m hcl at 110 c; 12-36 hours. Each peptide bond is broken and products are all of the free amino acids. Each amino acid is separated, identified and quantified. Final result: know how many of each amino acid present in the original. How to determine the order: determine the c-terminal amino acid. Use carboxypeptidase enzyme that removes the last (c-terminal) amino acid in a free form by breaking the peptide bond: hydrolyzes the peptide bond nearest the c-terminus, identify the n-terminal amino acids in order. Often difficult to characterize an intact protein. Instead, employ a divide and conquer approach to analyze peptide fragments of the intact protein. Use enzymes called proteases: cleave peptide bond in a specific way, two examples, trypsin cleaves on the c-terminal side of lys and arg residues, chymotrypsin cleaves on the c-terminal side of tyr, phe, and trp.