Biochemical Studies on Recombinant Human Isobutyryl-CoA Dehydrogenase
Nasser E. Ibrahim and Al-Walid A. Mohsen
DOI : 10.3844/ajbbsp.2011.84.89
American Journal of Biochemistry and Biotechnology
Volume 7, Issue 2
Problem statement: Human Isobutyryl-CoA Dehydrogenase (IBD) is member of the Acyl-CoA Dehydrogenases family. It is involved in Val metabolism. In this study we modified the purification method of the IBD and further spectroscopic characterization was conducted. Approach: Using DTT in the buffers during the purification, IBD purified to homogeneity by different chromatographic steps. Reactivity of Cys residues were measured using thiol modifying agent DNTB. Results: IBD found to be homotetramer with 42.7 KDa for each subunit and the value of its pI was 6.2. Conclusion: Thiol modifying reagent showed crucial role (s) of some Cys residues for IBD structure, FAD binding and/or activity.
© 2011 Nasser E. Ibrahim and Al-Walid A. Mohsen. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.