Date of Award

5-19-2011

Document Type

Thesis

Degree Name

Bachelor of Arts

First Advisor

Hofferberth, John

Abstract

The E. coli maltose-binding protein (MBP) is a robust binding protein with high affinity to maltose and a number of related polysaccharides. The combination of its stability and ability to bind a number of structurally similar ligands suggests that it may be useful as a scaffold protein that could be engineered to accept alternative ligands. We describe here our efforts to develop an efficient means of producing two MBP variants with engineering at the C-terminus: MBP-41 and MBP-6His. Further, a novel fluorescence assay to measure the binding affinity of MBP and its mutants to both fluorescently-labeled and unlabeled ligands is described. Preliminary equilibrium binding studies suggest that MBP-41 has similar affinity to maltose (KD 0.61 å± 0.16 ë_M) as wild-type MBP (KD 1.2 å± 0.20 ë_M). 1

Comments

Includes bibliographical references: pages 82-84

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