Bailey, Aaron

Ernat, Justin

Moore, Faculty Advisor, Laura

Digital Commons @ Illinois Wesleyan University

Illinois Wesleyan UniversityDigital Commons @ IWUJohn Wesley Powell Student ResearchConference 2006, 17th Annual JWP ConferenceApr 8th, 9:00 AM - 10:00 AMThe Analysis of Amino Acid Substitutions atPositions ASP22 of FNRJustin ErnatIllinois Wesleyan UniversityAaron BaileyIllinois Wesleyan UniversityLaura Moore, Faculty AdvisorIllinois Wesleyan UniversityFollow this and additional works at: http://digitalcommons.iwu.edu/jwprcThis Event is brought to you for free and open access by The Ames Library, the Andrew W. Mellon Center for Curricular and FacultyDevelopment, the Office of the Provost and the Office of the President. It has been accepted for inclusion in Digital Commons @ IWU bythe faculty at Illinois Wesleyan University. For more information, please contact digitalcommons@iwu.edu.©Copyright is owned by the author of this document.Justin Ernat; Aaron Bailey; and Laura Moore, Faculty Advisor, "The Analysis of Amino Acid Substitutions at PositionsASP22 of FNR" (April 8, 2006). John Wesley Powell Student Research Conference. Paper 1.http://digitalcommons.iwu.edu/jwprc/2006/posters/1THE .TOHN WESLEY POWELL STUDENT RESEARCH CONFERENCE - APRIL 2006 Poster Presentation P3 THE ANALYSIS OF AMINO ACID SUBSTITUTIONS AT POSITIONS ASP22 OF FNR Justin Emat and Aaron Bailey and Laura Moore* Chemistry Department, Illinois Wesleyan University The FNR protein is an oxygen-sensing transcription factor in the facultative anaerobe, Escherichia coli. In wild-type FNR, a [4Fe-4S] cluster in incorporated into the protein. This results in a protein that dimerizes and can bind to specific sites in the DNA. In the presence of oxygen, this cluster degrades causing a conformational change that renders the protein inactive. The Fe-S cluster is ligated by cystein residues at positions 20, 23, 29, and 1 22. Our work focuses on the effects on protein activity of amino acid substitutions at position Asp22. We studied the aerobic and anaerobic activity of the mutations Asp22Ala, Asp22Ser, Asp22Trp, Asp22Asn and Asp22Gly using Beta­galactosidase assays. Of these mutations Asp22Gly has been previously reported to be aerobically active, and it was suggested that this substitution stabilized the cluster to oxygen. Currently, under the low protein concentrations of our Beta-galactosidase assays, none substitutions have shown aerobic activity. We have also isolated Asp22Ala and Asp22G1y FNR protein. The Asp22Ala FNR protein has been characterized using absorption spectroscopy, which shows that the [4Fe-4S] cluster is not stable to the presence of oxygen. Further research will include isolation of other Asp22 FNR proteins as well as performing Beta-galactosidase assays with a higher concentration of protein. 

The Analysis of Amino Acid Substitutions at Positions ASP22 of FNR

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The Analysis of Amino Acid Substitutions at Positions ASP22 of FNR

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