Further study on the two pivotal parts of Hlg2 for the full hemolytic activity of staphylococcal gamma-hemolysin.
Staphylococcal gamma-hemolysin consists of LukF of 34 kDa and Hlg2 (or H gamma II) of 32 kDa, which cooperatively lyse human and rabbit erythrocytes. Our previous data showed that the 5-residue segment K23R24L25A26I27 of Hlg2 is pivotal for the hemolytic activity [Nariya, H. and Kamio, Y., Biosci. Biotechnol. Biochem., 59, 1603-1604 (1997)]. Here, we identify an additional amino acid residue in Hlg2 necessary for the full gamma-hemolysin activity by measuring the toxin activity of Hlg2 mutants in the presence of LukF. The data obtained showed that Arg217 of Hlg2 is an additional pivotal amino acid residue besides the KRLAI segment for the full Hlg2-specific function in gamma-hemolysin. We also report evidence that the Hlg2 mutants showing a low or null hemolytic activity in the presence of LukF towards human erythrocytes had low or no binding activity to the cells, resulting in failure of formation of the ring-shaped pore-forming complex on the erythrocytes.
Yokota
K
K
Department of Applied Microbiology, Graduate School of Agricultural Science, Tohoku University, Sendai, Japan.
Sugawara
N
N
Nariya
H
H
Kaneko
J
J
Tomita
T
T
Kamio
Y
Y
D000818:Animals; D001426:Bacterial Proteins; D001427:Bacterial Toxins / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D004912:Erythrocytes / Q000378:metabolism; D006460:Hemolysin Proteins / Q000378:metabolism*; D006461:Hemolysis; D006801:Humans; D007956:Leukocidins / Q000378:metabolism*; D011817:Rabbits; D013211:Staphylococcus aureus / Q000378:metabolism*
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; D007956:Leukocidins; C037956:gamma-hemolysin, Staphylococcus aureus; C074300:LukF protein, Staphylococcus aureus