MBB 222 Lecture Notes - Lecture 10: Protein Tertiary Structure, Christian B. Anfinsen, Guanidinium Chloride

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Tertiary structure: domains, globular proteins, protein folding and stability, covalent modification, quaternary structure. Disruption of the native structure (by breaking the noncovalent bonds responsible for secondary (2o) and tertiary (3o) structures) is called denaturation the result of denaturation is a protein in a non-native or denatured state. Denaturation can be caused by: raising the temperature: extremes of ph, (cid:272)haotropi(cid:272) age(cid:374)ts (cid:894)(cid:862)de(cid:374)atura(cid:374)ts(cid:863), like 8 m urea or 6 m gua(cid:374)idi(cid:374)iu(cid:373) (cid:272)hloride(cid:895, detergents that disrupt hydrophobic (and sometimes also polar interactions), etc. Most proteins are insoluble when denatured due to newly-exposed hydrophobic regions that cause protein aggregation and precipitation. Some proteins can refold after denaturation under the right conditions. Reduction of disulfide bonds can destabilize a protein and contribute to protein denaturation. Reducing agents like b-mercaptoethanol (b-me) and dithiothreitol (dtt) reduce disulfide bonds: as the disulfide bonds are reduced, the reducing agent is oxidized and forms disulfide- bonded dimers.

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