Date of Award

2006

Degree Type

Thesis

Degree Name

Master of Science

Program

Biochemistry

Supervisor

Dr. Eric H. Ball

Second Advisor

Dr. David Edgell

Third Advisor

Dr. David Litchfield

Abstract

Vinculin and metavinculin are components of cell-matrix and cell-cell adherens junctions, differing in a 68 residue insert in the metavinculin C-terminus. These proteins are known to interact with the EVHI domain of VASP through a proline-rich motif. A second probable proline-rich motif was identified in the metavinculin insert. The intent of this study was to determine whether a second motif in the metavinculin insert was a VASP binding site. A number of solution phase assays proved unable to detect an interaction through either site, but a solid-phase, overlay assay could do so. Mutations in the two motifs were constructed and VASP binding measured. Both motifs were found to contribute to the binding. A fluorescent probe was able to restore binding to both motifs in place of the native phenylalanine. Thus, it appears that the insert proline-rich motif may be a VASP binding site.

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