Characterization of a haemolytic factor from Candida albicans.
The culture supernatant of Candida albicans promoted the disruption of human red blood cells (RBCs). The haemolytic activity was detected in a sugar-rich fraction (about 200 kDa) from Sephacryl S-100 chromatography. As the haemolytic activity was adsorbed by concanavalin A-Sepharose, the haemolytic factor may be a mannoprotein. The activity was inactivated by periodate oxidation, indicating that the sugar moiety of the mannoprotein played an important role in the haemolysis. The structure of the sugar moiety of the mannoprotein was identified as a cell-wall mannan by 1H-NMR analysis, and purified C. albicans mannan promoted the disruption of RBCs. The binding of mannan to RBCs was demonstrated by flow cytometric analysis and was inhibited by the addition of band 3 protein inhibitor, 4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid (DIDS). The haemolysis caused by mannan was inhibited by DIDS, SITS (4-acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic acid) and bis(sulfosuccinimidyl) suberate, but not by pyridoxal 5-phosphate. These results indicated that a mannoprotein released from C. albicans bound to the band 3 protein on RBCs, thereby promoting their disruption.
Watanabe
Toshihiko
T
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Takano
Mari
M
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Murakami
Mariko
M
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Tanaka
Hironori
H
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Matsuhisa
Atsushi
A
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Nakao
Noriko
N
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Mikami
Takeshi
T
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
Suzuki
Masuko
M
Sendai Research Institute for Mycology, 14-34 Toshogu 1 Chome, Aobaku, Sendai 981-0908, Japan.
Matsumoto
Tatsuji
T
Department of Microbiology, Tohoku College of Pharmacy, 4-1 Komatsushima 4 Chome, Aoba-ku, Sendai 981-8558, Japan.
D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid / Q000494:pharmacology; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid / Q000494:pharmacology; D000838:Anions / Q000378:metabolism; D001692:Biological Transport / Q000187:drug effects; D002176:Candida albicans / Q000737:chemistry*; D002473:Cell Wall / Q000737:chemistry; D004912:Erythrocytes / Q000378:metabolism; D006461:Hemolysis / Q000187:drug effects; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy; D008351:Mannans / Q000302:isolation & purification* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000302:isolation & purification* / Q000378:metabolism; D013388:Succinimides / Q000494:pharmacology
D000838:Anions; D008351:Mannans; D008562:Membrane Glycoproteins; D013388:Succinimides; C025363:mannoproteins; D012856:4-Acetamido-4'-isothiocyanatostilbene-2,2'-disulfonic Acid; D006859:Hydrogen; C035760:bis(sulfosuccinimidyl)suberate; D017878:4,4'-Diisothiocyanostilbene-2,2'-Disulfonic Acid