Abstract
Many species of gram-positive bacteria produce branched peptidoglycan precursors resulting from the transfer of various L-amino acids or glycine from amino acyl-tRNA to the epsilon-amino group of L-lysine. The UDP-MurNAc-pentapeptide:L-alanine ligase and alanyl-tRNA synthetase genes from Enterococcus faecalis were identified, cloned, and overexpressed in Escherichia coli. The purified enzymes were necessary and sufficient for tRNA-dependent addition of L-alanine to UDP-MurNAc-pentapeptide in vitro. The ligase belonged to the Fem family of proteins, which were initially identified genetically as factors essential for methicillin resistance in Staphylococcus aureus.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Alanine / metabolism
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Alanine-tRNA Ligase / metabolism*
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Chromatography, High Pressure Liquid
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Enterococcus faecalis / enzymology
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Enterococcus faecalis / genetics
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Enterococcus faecalis / metabolism*
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Escherichia coli / enzymology
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Escherichia coli / genetics
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Lactobacillaceae / enzymology
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Lactobacillaceae / genetics
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Mass Spectrometry
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Methicillin Resistance / genetics
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Peptidoglycan / biosynthesis*
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Protein Precursors / biosynthesis*
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Staphylococcus aureus / genetics
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Uridine Diphosphate N-Acetylmuramic Acid / analogs & derivatives
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Uridine Diphosphate N-Acetylmuramic Acid / metabolism*
Substances
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Peptidoglycan
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Protein Precursors
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Uridine Diphosphate N-Acetylmuramic Acid
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UDP-N-acetylmuramic acid pentapeptide
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Alanine-tRNA Ligase
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Alanine