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Glyoxalase 2-2 over-expression and characterization of a metallohydrolase from Arabidopsis thaliana /

by Wenzel, Nathan F.

Abstract (Summary)
GLYOXALASE 2-2: OVER-EXPRESSION AND CHARACTERIZATION OF A METALLOHYDROLASE FROM ARABIDOPSIS THALIANA By Nathan F. Wenzel In order to investigate the metal binding properties of A. thaliana glyoxalase 2-2, recombinant GLX 2-2 was over-expressed in rich or minimal media containing various amounts of added Zn(II), Fe(II), or Mn(II). The resulting enzymes were purified, characterized, and compared. Kinetic studies and metal analyses of the samples indicated that no significant catalytic differences existed between the enzymes, in spite of the enzymes binding different types and amounts of metal ions. EPR and EXAFS spectroscopies strongly indicated the presence of positive cooperativity of metal binding and of numerous possible dinuclear metal centers, including Fe(III)Zn(II), Fe(III)Fe(II), and Mn(II)Mn(II) centers. The ability to bind various amounts of manganese, iron, and zinc with no apparent change in catalytic activity and the possibility of positive cooperativity in metal binding make A. thaliana glyoxalase 2-2 unique to the family of proteins containing the ?-lactamase fold. Glyoxalase 2-2: Over-expression and Characterization of a Metallohydrolase from Arabidopsis thaliana A Thesis Submitted to the Faculty of Miami University In partial fulfillment of The requirement for the degree of Master of Science Department of Chemistry and Biochemistry By Nathan F. Wenzel Miami University Oxford, OH 2003 Advisor________________________ Michael W. Crowder Reader_________________________ Christopher A. Makaroff Reader_________________________ John W. Hawes Reader_________________________ Robert E. Minto
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Advisor:

School:Miami University

School Location:USA - Ohio

Source Type:Master's Thesis

Keywords:glyoxalase binding sites biochemistry

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