CHARACTERIZATION OF THE COMPONENTS OF HEMOLYSIN-BL FROM BACILLUS-CEREUS
CHARACTERIZATION OF THE COMPONENTS OF HEMOLYSIN-BL FROM BACILLUS-CEREUS
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DOI：
10.1128/iai.59.5.1778-1784.1991
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发表时间：
1991-05-01
影响因子：
3.1
通讯作者：
MACMILLAN, JD
中科院分区：
文献类型：
--
作者：
BEECHER, DJ;MACMILLAN, JD
在此之前，我们对蜡状芽孢杆菌中一种新的溶血素进行了部分纯化，结果表明，溶血活性需要至少两个组分B和L的共同作用，以表明它们在这一活性中的细胞结合和细胞溶解作用。在进一步纯化时，如本文所述，结合阴离子交换层析和聚丙烯酰胺凝胶电泳法分离出三种蛋白质B、L1和L2(分别为35、36和45 kDa)。单独地，这些蛋白在溶血和血管通透性分析中是无效的，而B和L1或B和L2的组合要么不活性，要么略有活性。这三个部分的结合产生了独特的环状溶血区，这是溶血素BL的一个特征，以及血管通透性分析中的浮肿和发蓝。由于已知的血管通透性测定与肠毒性有关，这些结果表明溶血素BL是肠毒素。此外，蜡样芽胞杆菌多组分腹泻性肠毒素的溶血素BL组分的分子量和等电点与其他人所描述的一致。经B处理的红细胞的免疫荧光染色证实，在L成分能够引起细胞溶解之前，B与细胞结合是所需的第一步。
Previously we described the partial purification of a novel hemolysin from Bacillus cereus and showed that hemolytic activity required the combined action of at least two components, called B and L to signify their cell-binding and cell-lytic roles in this activity. On further purification, as described in the present article, a combination of anion-exchange chromatography and polyacrylamide gel electrophoresis separated three proteins, B, L1, and L2 (35, 36, and 45 kDa, respectively). Individually, these proteins were inactive in hemolytic and vascular permeability assays, and combinations of B and L1 or B and L2 were either inactive or slightly active. Combinations of all three moieties produced the unique ring-shaped zone of hemolysis, a previously described characteristic of hemolysin BL, as well as edema and bluing in the vascular permeability assay. Since the vascular permeability assay is known to correlate with enterotoxicity, these results suggest that hemolysin BL is enterotoxigenic. Furthermore, the molecular weights and isoelectric point values of the hemolysin BL components are consistent with those described by others for the multicomponent diarrheal enterotoxin of B. cereus. Immunofluorescent staining of B-treated erythrocytes confirmed that B binds to cells as an initial step required before the L components can act to cause cell lysis.