BCH 2333 Lecture Notes - Lecture 11: Edman Degradation, Mass Spectrometry, Protein Purification

• School: University of Ottawa
• Department: Biochemistry
• Course: BCH 2333
• Professor: Matthew Lafreniere

Document Summary

Good for protein that you want to study. Primary sequence of a polypeptide will provide info on: We can compare primary sequences with other biological molecules. Successive rounds of n-terminal modification, cleavage, and identification. Can be used to identify protein with known sequence. Used to se(cid:395)ue(cid:374)(cid:272)e (cid:396)i(cid:271)o(cid:374)u(cid:272)lease (cid:894)(cid:1005)(cid:1006)(cid:1008) a. a. (cid:895) i(cid:374) (cid:1005)9(cid:1009)(cid:1004)s a(cid:374)d -galactosidase (1021 a. a. ) in 1978. Need to go through a bunch of sequences so it"s super slow and no one uses it anymore. Mass spectrometry (modern method) - used all the time. Maldi m a(cid:374)d e i m (cid:272)a(cid:374) (cid:271)e p(cid:396)e(cid:272)isely ide(cid:374)tify the (cid:373)ass of a peptide a(cid:374)d it"s a(cid:373)i(cid:374)o a(cid:272)id sequence. Each polypeptide has a particular mass spec finger print. Get ride of the disulfide bonds -> you want a linear polypeptide when you are seqeuncing. These bonds hold tertiary and quaternary structures together. The green one is to cap the open thiols. Makes sure it doesn"t oxidise to form thiols again.