Cloning, sequencing, and expression of the adenosylcobalamin-dependent ribonucleotide reductase from Lactobacillus leichmannii

S. Booker, J. Stubbe

Research output: Contribution to journalArticle

91 Scopus citations

Abstract

Ribonucleoside-triphosphate reductase (RTPR, EC 1.17.4.2) from Lactobacillus leichmannii, a monomeric adenosylcobalamin-requiring enzyme, catalyzes the conversion of nucleoside triphosphates to deoxynucleoside triphosphates. The gene for this enzyme has been cloned and sequenced. In contrast to expectations based on mechanistic considerations, there is no statistically significant sequence homology with the Escherichia coli reductase that requires a dinuclear-iron center and tyrosyl radical cofactor. The RTPR has been overexpressed and purified to homogeneity, yielding 90 mg of protein from 2.5 g of bacteria. Initial characterization of the recombinant RTPR indicates that its properties are identical to those of the RTPR isolated from L. leichmannii.

Original languageEnglish (US)
Pages (from-to)8352-8356
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume90
Issue number18
DOIs
StatePublished - 1993

All Science Journal Classification (ASJC) codes

  • General

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