In vitro phosphinate methylation by PhpK from Kitasatospora phosalacinea.

@article{Werner2011InVP,
  title={In vitro phosphinate methylation by PhpK from Kitasatospora phosalacinea.},
  author={W. J. Werner and Kylie D. Allen and K. Hu and G. Helms and Brian Chen and Susan C Wang},
  journal={Biochemistry},
  year={2011},
  volume={50 42},
  pages={
          8986-8
        }
}
Radical S-adenosyl-L-methionine, cobalamin-dependent methyltransferases have been proposed to catalyze the methylations of unreactive carbon or phosphorus atoms in antibiotic biosynthetic pathways. To date, none of these enzymes has been purified or shown to be active in vitro. Here we demonstrate the activity of the P-methyltransferase enzyme, PhpK, from the phosalacine producer Kitasatospora phosalacinea. PhpK catalyzes the transfer of a methyl group from methylcobalamin to 2-acetylamino-4… Expand
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References

SHOWING 1-10 OF 30 REFERENCES
A Radically Different Mechanism for S-Adenosylmethionine–Dependent Methyltransferases
TLDR
The mechanisms of two reactions targeting the sp2-hybridized carbons at positions 2 and 8 of adenosine 2503 in 23S ribosomal RNA, catalyzed by RlmN and Cfr, respectively are determined. Expand
Product inhibition in the radical S‐adenosylmethionine family
TLDR
It is demonstrated that three members of the radical S‐adenosylmethionine superfamily are inhibited in vitro by a combination of the products 5′‐deoxyadenosine (DOA) and methionine. Expand
RNA methylation by Radical SAM enzymes RlmN and Cfr proceeds via methylene transfer and hydride shift
TLDR
Findings indicate that rather than acting as methyltransferases, RlmN and Cfr are methyl synthases, and demonstrate that all three carbon atoms attached to the sulfonium center in SAM can serve as precursors to carbon-derived radicals in enzymatic reactions. Expand
Sequence of a P-methyltransferase-encoding gene isolated from a bialaphos-producing Streptomyces hygroscopicus.
TLDR
The amino-acid sequence deduced from the nt sequence, shows homology with those of magnesium-protoporphyrin IX monomethyl ester oxidative cyclase (Mg-ProtoMe cyclase) of Rhodobacter capsulatus and the enzyme catalyzing the methylation of the aldehyde carbon of phosphonoacetaldehyde in fosfomycin biosynthesis. Expand
Nucleotide sequence of fortimicin KL1 methyltransferase gene isolated from Micromonospora olivasterospora, and comparison of its deduced amino acid sequence with those of methyltransferases involved in the biosynthesis of bialaphos and fosfomycin.
TLDR
PnAA was directly methylated by a nucleophilic attack of the methyl anion derived from methylcobalamin catalyzed by PnAA methyltransferase, and the nucleotide sequences of the genes encoding P-methyltransferase were determined. Expand
Studies on the biosynthesis of bialaphos (SF-1293) 12. C-P bond formation mechanism of bialaphos: discovery of a P-methylation enzyme.
An enzymatic activity catalyzing P-methylation of N-acetyldemethylphosphinothricin, a biosynthetic intermediate of the herbicide bialaphos, was detected in a cell extract of StreptomycesExpand
Biotin synthase exhibits burst kinetics and multiple turnovers in the absence of inhibition by products and product-related biomolecules.
TLDR
This work confirms that dAH is a modest inhibitor of BS and shows that cooperative binding of dAH with excess methionine results in a 3-fold enhancement of this inhibition, and demonstrates that AdoHcy is a potent inhibitors of BS while MTA is not an inhibitor. Expand
Studies on the biosynthesis of bialaphos (SF-1293). 2. Isolation of the first natural products with a C-P-H bond and their involvement in the C-P-C bond formation.
TLDR
2-Phosphinomethylmalic acid synthase catalyzes the condensation of phosphinopyruvic acid, an analog of oxalacetic acid, and acetyl-CoA to form PMM, which is very similar to (R)-citrate synthase of Clostridium in the inhibition pattern by sulfhydryl compounds, its metal ion requirement and stereospecificity. Expand
Anaerobic functionalization of unactivated C-H bonds.
  • S. Booker
  • Chemistry, Medicine
  • Current opinion in chemical biology
  • 2009
TLDR
This review will focus on a special subclass of radical SAM enzymes that functionalize inert C-H bonds, highlighting the functional groups and the chemistry that leads to their insertion. Expand
Studies on the biosynthesis of bialaphos (SF-1293). 1. Incorporation of 13C- and 2H-labeled precursors into bialaphos.
TLDR
The biosynthetic studies on I, a tripeptide comprising two alanine residues and one phosphinothricin, showed an AB-type 13C-13C coupling between C-4 and C-3 providing unequivocal evidence that these two carbon atoms in II originate from the intact acetic acid molecule. Expand
...
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