Characterization of the Staphylococcus aureus mprF gene, involved in lysinylation of phosphatidylglycerol.
Lysylphosphatidylglycerol (LPG) is a basic phospholipid in which L-lysine from lysyl-tRNA is transferred to phosphatidylglycerol (PG). This study examined whether the Staphylococcus aureus mprF gene encodes LPG synthetase. A crude membrane fraction prepared from wild-type S. aureus cells had LPG synthetase activity that depended on PG and lysyl-tRNA, whereas the membrane fraction from an mprF deletion mutant did not. When S. aureus MprF protein was trans-expressed in wild-type Escherichia coli cells, LPG synthesis was induced, whereas it was not observed in E. coli pgsA3 mutant cells in which the amount of PG is significantly reduced. In addition, LPG synthetase activity and a 93 kDa protein whose molecular size corresponded to that of MprF protein were co-induced in the crude membrane fraction prepared from E. coli cells expressing MprF protein. The Km values of the LPG synthetase activity for PG and for lysyl-tRNA were 56 microM and 6.9 microM, respectively, consistent with those of S. aureus membranes. These results suggest that the MprF protein is LPG synthetase.
Oku
Yusuke
Y
Laboratory of Developmental Biochemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 3-1 7-Chome, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Kurokawa
Kenji
K
Laboratory of Developmental Biochemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 3-1 7-Chome, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Ichihashi
Norikazu
N
Laboratory of Developmental Biochemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 3-1 7-Chome, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
Sekimizu
Kazuhisa
K
Laboratory of Developmental Biochemistry, Graduate School of Pharmaceutical Sciences, The University of Tokyo, 3-1 7-Chome, Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
D019881:Aminoacyltransferases; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D005798:Genes, Bacterial; D007700:Kinetics; D008239:Lysine; D008246:Lysophospholipids / Q000096:biosynthesis*; D010715:Phosphatidylglycerols / Q000378:metabolism; D012346:RNA, Transfer, Amino Acyl / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D013211:Staphylococcus aureus / Q000235:genetics* / Q000378:metabolism*
D001426:Bacterial Proteins; D008246:Lysophospholipids; D010715:Phosphatidylglycerols; D012346:RNA, Transfer, Amino Acyl; D011994:Recombinant Proteins; C002285:lysylphosphatidylglycerol; D019881:Aminoacyltransferases; C429146:mprF protein, Staphylococcus aureus; D008239:Lysine