CUP Undergraduate Research

Title

X-Ray Crystallographic Analysis of Uncharacterized Bacillus cereus Protein Reveals Nucleotide Binding

Date of Award

Spring 4-1-2016

Document Type

Restricted Access Thesis

College

College of Theology, Arts, & Sciences

Department

Math & Science

Degree Name

Biology, BA

First Advisor

Michael H. Godsey, PhD

Abstract

Bacillus cereus is a soil born opportunistic pathogen known to act as a key player in food poisoning (Bottone, 2010). Its close relation to Bacillus anthracis makes it a protein of interests for biochemists. Protein BC2332 is a unique protein within Bacillus cereus with limited structural similarity to ADP – ribosylating toxins. This particular protein class, ADPribosylating toxins, has a common function in that they bind to NADH (Todar, 2012). To prove that BC2332 belongs in the class of ADP – ribosylating toxins it is necessary to prove that BC2332 binds with NADH. The Development of a successful buffer for the protein BC2332 from Bacillus cereus allowed for purification of this uncharacterized protein. Successful Crystallographic and X-ray analysis reveal density for putative nucleotide binding in the active site of BC2332, with implications for a possible function as an ADP – ribosylating toxin. Electron density of BC2332 in crystalline structure from the x-ray data, reveled density in the putative active site which is consistent with NADH binding similarly with other ADP-ribosylating toxins.

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