Synthesis of aromatic monothiols to increase the folding rate and yields of disulfide-containing proteins

Document Type

Thesis

Degree

Master of Science (MS)

Major/Program

Chemistry

First Advisor's Name

Watson Lees

First Advisor's Committee Title

Committee Chair

Second Advisor's Name

David Chatfield

Third Advisor's Name

J. Martin E. Quirke

Date of Defense

7-15-2011

Abstract

Almost all pharmaceutically relevant proteins and many extracellular proteins contain disulfide bonds, which are essential for protein functions. In many cases, disulfidecontaining proteins are produced via in vitro protein folding that involves the oxidation of reduced protein to native protein, a complex process. The in vitro folding of reduced lysozyme has been extensively studied as a model system because native lysozyme is small, inexpensive, and has only four disulfide bonds. The folding of reduced lysozyme is conducted with the aid of a redox buffer consisting of a small molecule disulfide and a small molecule thiol, such as oxidized and reduced glutathione. Herein, in vitro folding rates and yields of lysozyme obtained in the presence of a series of aromatic thiols and oxidized glutathione are compared to those obtained with reduced and oxidized glutathione. Results showed that aromatic thiols significantly increase the folding rate of lysozyme compared to glutathione.

Identifier

FI14050441

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