Date of Award
1997
Publication Type
Master Thesis
Degree Name
M.Sc.
Department
Chemistry and Biochemistry
Keywords
Chemistry, Biochemistry.
Supervisor
Szabo, A. G.,
Rights
info:eu-repo/semantics/openAccess
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-No Derivative Works 4.0 International License.
Abstract
An M13 Phage Display library experiment by Dr. B. Kay (University of North Carolina) yielded a lone Ca$\sp{2+}$ dependent calmodulin-binding peptide (12.1). Point mutations to the C-terminus and the central Gly 13 Gly 14 residues of this peptide were made to study their effect on the peptide binding to Cam. This series of peptides was denoted as the 'Evolved' series of peptides (52). Two sources of this peptide series were used during experiments, one provided by Dr. B. Kay, University of North Carolina (K series) and a second from Dr. G. Laloie, University of Waterloo (UW series). Fluorescence spectral titrations with Cam indicated that the results were source dependent. Secondly, fluorescence titrations indicated that the peptides bound to Cam in 1:1 or 2:1 equivalents of peptide:Cam. Scatchard analysis enabled the K$\sb{\rm d}$'s one or both complexes to be determined. Comparison of the K$\sb{\rm d}$'s obtained, (ranging from $$ E3-AS $>$ E5 $>$ E4 $>$ E6 $>$ E3-LTV $>$ 12.1. A second project studied the binding interactions between the peptide analogues of sm-MLCK, neuromodulin and the Ras-like protein KirGem. Data for sm-MLCK and KirGem indicated 2:1 binding. Scatchard analysis estimated the K$\sb{\rm d}$'s for sm-MLCK as 0.56 $\mu$M for the 1:1 complex and $\sim$16 $\mu$M for the 2:1 complex and the K$\sb{\rm d}$ of the 2:1 KirGem complex as 7.2 $\mu$M. Studies with neuromodulin indicated stoichiometric binding and further that Tyr 138 of Cam participates in binding while Tyr 99 does not. (Abstract shortened by UMI.)Dept. of Chemistry and Biochemistry. Paper copy at Leddy Library: Theses & Major Papers - Basement, West Bldg. / Call Number: Thesis1997 .S73. Source: Masters Abstracts International, Volume: 37-02, page: 0616. Adviser: A. G. Szabo. Thesis (M.Sc.)--University of Windsor (Canada), 1997.
Recommended Citation
Stevenson, Wendy Joan., "Characterization of calmodulin binding properties of a phage display peptide series and the peptide analogues of KirGem sm-MLCK and neuromodulin using fluorescence spectroscopy." (1997). Electronic Theses and Dissertations. 1397.
https://scholar.uwindsor.ca/etd/1397