Senior Project Advisor

Anthony-Cahill, Spencer J.

Document Type

Project

Publication Date

Spring 2001

Keywords

Permuteins

Abstract

Our research is aimed at generating and characterizing topological mutants of sperm whale myoglobin (swMb). The long-term experimental design involves the production of circular permuteins from swMb genes fused by a linker and determination of the effect of linker sequence on protein stability. We have expressed circularly permuted sperm whale myoglobins in E. coli including variants that start at the C helix and end with the B helix (CBLx), and others beginning at the H helix and ending with the G helix (HGLx). Expression yields for mutant myoglobins have been shown to correlate with stabilities of the mutants (Hargrove et ai. Biochemistry, 1994, 33, 11767). We have observed reduced levels of expression for our permuted myoglobin mutants. In order to improve the expression yields for destabilized topological mutants of myoglobin, we are attempting the expression of Glutathione S-transferase (GST)-permutein fusion proteins. The GST fusion system has been shown to be useful in expression, purification, and detection of proteins (Smith, Methods Mol Cell Biol 1993, 4, 220). We hope this will provide a general strategy for increased expression of our permuteins.

Department

Chemistry

Subjects - Topical (LCSH)

Proteins--Chemical modification; Myoglobin; Sperm whale

Genre/Form

student projects; term papers

Type

Text

Rights

Copying of this document in whole or in part is allowable only for scholarly purposes. It is understood, however, that any copying or publication of this document for commercial purposes, or for financial gain, shall not be allowed without the author’s written permission.

Rights Statement

http://rightsstatements.org/vocab/InC/1.0/

Language

English

Format

application/pdf

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Included in

Chemistry Commons

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