期刊
ARCHIVES OF BIOCHEMISTRY AND BIOPHYSICS
卷 459, 期 1, 页码 1-9出版社
ELSEVIER SCIENCE INC
DOI: 10.1016/j.abb.2006.12.024
关键词
pyruvate formate-lyase PFL; AdhE; glycyl radical quenching; deactivation of glycyl radical enzyme
资金
- NIGMS NIH HHS [GM54608, R29 GM054608, R01 GM054608] Funding Source: Medline
Escherichia coli AdhE has been reported to harbor three distinct enzymatic activities: alcohol dehydrogenase, acetaldehyde-CoA dehydrogenase, and pyruvate formate-lyase (PFL) deactivase. Herein we report on the cloning, expression, and purification of E. coli AdhE, and the re-investigation of its purported enzymatic activities. While both the alcohol dehydrogenase and acetaldehyde-CoA dehydrogenase activities were readily detectable, we were unable to obtain any evidence for catalytic deactivation of PFL by AdhE, regardless of whether the reported cofactors for deactivation (Fe(II), NAD, and CoA) were present. Our results demonstrate that AdhE is not a PFL deactivating enzyme. We have also examined the potential for deactivation of active PFL by small-molecule thiols. Both beta-mercaptoethanol and dithiothreitol deactivate PFL efficiently, with the former providing quite rapid deactivation. PFL deactivated by these thiols can be reactivated, suggesting that this deactivation is non-destructive transfer of an H atom equivalent to quench the glycyl radical. (c) 2007 Elsevier Inc. All rights reserved.
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