Masters Theses

Date of Award

8-2000

Degree Type

Thesis

Degree Name

Master of Science

Major

Biochemistry and Cellular and Molecular Biology

Major Professor

Engin H. Serpersu

Committee Members

Cynthie B. Peterson

Abstract

Aminoglycoside nucleotidyltransferase (2")-la (ANT) was cloned from P Aeruginosa and purified from overexpressing £ coli BL21 (DE3) cells. The high yields obtainable from this system will permit in-depth structural studies in the future. In addition, the conformation of the aminoglycoside antibiotic isepamicin, a psuedo-tri-saccharide, bound to aminoglycoside nucleotidyltransferase 2"-la has been determined using NMR spectroscopy. Molecular modeling employing experimentally determined inter-proton distances resulted in two different enzyme-bound conformations (conformer 1 and conformer 2) of isepamicin when comparing the structures using the 2-deoxystreptamine ring (B ring) and the D-garosamine ring (0 ring) Conformer 1 was by far the major conformer defined by the average glycosidic dihedral angles (Pec -65.26 ± 1 63 and '+'bc-54 76 ± 464 (20 structures). Conformer 1 was further subdivided into one major (conformer 1a)and two minor components (conformer 1 b and 1c) based on the comparison of O^b and Conformer 1a and conformer 2 have almost identical

Download
Files over 3MB may be slow to open. For best results, right-click and select "save as..."

Share

COinS