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
Recommended Citation
Taylor, Margaret, "Development of a Robust Expression System for E. coli Maltose-Binding Protein and a Novel Fluorescence Assay to Characterize Ligand Binding" (2011). Honors Theses. 43.
https://digital.kenyon.edu/honorstheses/43
Rights Statement
All rights reserved. This copy is provided to the Kenyon Community solely for individual academic use. For any other use, please contact the copyright holder for permission.
Comments
Includes bibliographical references: pages 82-84