Purification, cloning and characterization of variant LukE-LukD with strong leukocidal activity of staphylococcal bi-component leukotoxin family.
Staphylococcus aureus produces bi-component leukotoxins composed of non-associated soluble proteins, S and F. Neither S nor F component alone is cytotoxic, but components together are active. These include Panton-Valentine leukocidin (PVL), gamma-hemolysin, LukE-LukD and others. Purification of leukotoxin from Staphylococcus aureus V8 strain (ATCC 27733) which does not have PVL genes, identified an F component with 100% identical to that of PVL in the first twenty-five N-terminal amino acids. Molecular cloning of this toxin obtained 2,595 nucleotides sequences containing two novel open reading frames for S and F. Deduced amino acid sequences of the S and F were respectively 91 and 94% identical to those of LukE and LukD. These were named variant of LukE-LukD (LukEv-LukDv). The activity of the recombinant LukEv-LukDv to rabbit leukocytes was similar to that of recombinant PVL. LukEv-LukDv was hemolytic to rabbit red blood cells although the activity was only 8% of gamma-hemolysin, but PVL was not. These activities were quite different from the LukE-LukD which was reported no hemolytic and poorly cytotoxic to leukocytes compared to PVL. The lukEv-IukDv was found in 87% of clinical isolates of Staphylococcus aureus but lukE-lukD was not detected. These data demonstrate the existence of variant LukE-LukD in V8 strain (ATCC 27733).
Morinaga
Naoko
N
Department of Molecular Infectiology, Graduate School of Medicine, Chiba University, Chiba, Chiba 260-8670, Japan. morinaga@med.m.chiba-u.ac.jp
Kaihou
Yuriko
Y
Noda
Masatoshi
M
D000595:Amino Acid Sequence; D000818:Animals; D001426:Bacterial Proteins; D001483:Base Sequence; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005098:Exotoxins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D007956:Leukocidins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism / Q000494:pharmacology; D007962:Leukocytes / Q000378:metabolism; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D011817:Rabbits; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D013211:Staphylococcus aureus / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005098:Exotoxins; D007956:Leukocidins; C115218:LukD protein, Staphylococcus aureus; C115217:LukE protein, Staphylococcus aureus; D011994:Recombinant Proteins