Identification of the UDP-MurNAc-pentapeptide:L-alanine ligase for synthesis of branched peptidoglycan precursors in Enterococcus faecalis.
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.
Bouhss
A
A
INSERM E0004-LRMA, UFR Broussais-Hôtel Dieu, Université Paris VI, 75270 Paris, France.
Josseaume
N
N
Allanic
D
D
Crouvoisier
M
M
Gutmann
L
L
Mainardi
J L
JL
Mengin-Lecreulx
D
D
van Heijenoort
J
J
Arthur
M
M
D000409:Alanine / Q000378:metabolism; D000412:Alanine-tRNA Ligase / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D013293:Enterococcus faecalis / Q000201:enzymology / Q000235:genetics / Q000378:metabolism*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D007777:Lactobacillaceae / Q000201:enzymology / Q000235:genetics; D013058:Mass Spectrometry; D016106:Methicillin Resistance / Q000235:genetics; D010457:Peptidoglycan / Q000096:biosynthesis*; D011498:Protein Precursors / Q000096:biosynthesis*; D013211:Staphylococcus aureus / Q000235:genetics; D014538:Uridine Diphosphate N-Acetylmuramic Acid / Q000031:analogs & derivatives / Q000378:metabolism*
D010457:Peptidoglycan; D011498:Protein Precursors; D014538:Uridine Diphosphate N-Acetylmuramic Acid; C002225:UDP-N-acetylmuramic acid pentapeptide; D000412:Alanine-tRNA Ligase; D000409:Alanine