Date of Award

Spring 1-1-2016

Document Type

Thesis

Degree Name

Master of Science (MS)

Department

Chemistry & Biochemistry

First Advisor

Marcelo C. Sousa

Second Advisor

Robert T. Batey

Third Advisor

James A. Goodrich

Abstract

In E. coli, the β-barrel Assembly Machinery (BAM) is an outer membrane protein complex which catalyzes the folding and insertion of integral outer membrane proteins. This thesis explores the hypothesis that BAM recognizes substrates by a sequence specific motif. A high-throughput screening approach called the SPOT assay is used to determine if BAM subunits bind to previously implicated sequence motifs or some yet unidentified consensus sequence. For this assay, solutions of fluorescently labeled BAM subunits are used to probe peptide arrays displaying sequences derived from BAM substrates, and binding is visualized by increased fluorescence. Results of these experiments do not support the original hypothesis of sequence-specific recognition by BAM, and we propose that BAM may recognize substrates in a general fashion that does not require a targeting sequence.

Included in

Biochemistry Commons

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