Adenosylcobalamin-Dependent Ribonucleoside Triphosphate Reductase fromLactobacillus Leichmannii. Rapid, Improved Purification Involving dGTP-Based Affinity Chromatography Plus Biophysical Characterization Studies Demonstrating Enhanced, “Crystallographic Level” Purity
- 1 January 1999
- journal article
- research article
- Published by Taylor & Francis in Preparative Biochemistry & Biotechnology
- Vol. 29 (3) , 273-309
- https://doi.org/10.1080/10826069908544929
Abstract
Ribonucleoside triphosphate reductase (RTPR, EC 1.17.4.2) from Lactobacillus leichmannii is a 5′-deoxyadenosylcobalamin-dependent (AdoCbl; Coenzyme B12) enzyme. RTPR is also a prototypical adenosylcobalamin-dependent ribonucleotide reductase, one that, as its name indicates, converts ribonucleoside triphosphates (NTP) to deoxyribonucleoside triphosphates (dNTP). Upon substrate binding to RTPR, AdoCbl's cobalt-carbon bond is cleaved to generate cob(II)alamin, 5′-deoxyadenosine, and the cysteine (C408) derived thiyl radical. Five key cysteines (Cys 119, 408, 419, 731, and 736), from among the ten total cysteines, are involved in RTPR's catalytic mechanism.Keywords
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