Comparative Biochemistry and Physiology B: Biochemistry & Molecular Biology ( IF 2.2 ) Pub Date : 2021-09-14 , DOI: 10.1016/j.cbpb.2021.110673 Marcelo Victorio De Los Santos 1 , José Luis Sánchez-Salgado 2 , Ali Pereyra 2 , Edgar Zenteno 2 , Norberto Vibanco-Pérez 3 , Gabriela Ramos-Clamont Montfort 4 , Sonia A Soto-Rodriguez 5
Vibrio parahaemolyticus toxin PirABvp is the major virulence factor exotoxin that contributes to the disruption of the hepatopancreatic epithelium in acute hepatopancreatic necrosis disease in shrimp. The PirBvp subunit is a lectin that recognizes amino sugars; however, its potential role in recognition of the hepatopancreas has not been identified. In the present work, we identified the cellular receptor for PirBvp in the shrimp hepatopancreas. A ligand blot assay of hepatopancreas lysate showed that the PirBvp subunit recognizes two glycoprotein bands of 60 and 70 kDa (Gc60 and Gc70). The hepatopancreas lysate was fractionated by anion-exchange chromatography, and the three main fractions obtained contained the recognized Gc60 and Gc70 protein bands. LC-MS/MS indicated that beta-hexosaminidases subunit beta and mucin-like 5 AC corresponded to the 60 and 70 kDa bands, respectively, which seem to be expressed in the epithelial cells of the hepatopancreas. Endoglycosidase treatment of hepatopancreas lysate with the O-glycosidase from Enterococcus faecalis, inhibits the binding of PirBvp. Altogether, these results suggest the relevance of the interaction of PirBvp with the hepatopancreas in the pathogenesis of acute hepatopancreatic necrosis disease in shrimp.
中文翻译:
副溶血性弧菌亚单位毒素 PirBvp 识别南美白对虾肝胰腺上皮上的糖蛋白
副溶血性弧菌毒素 PirAB vp是主要的毒力因子外毒素,在虾的急性肝胰腺坏死疾病中导致肝胰腺上皮细胞的破坏。PirB vp亚基是一种识别氨基糖的凝集素;然而,尚未确定其在识别肝胰腺方面的潜在作用。在目前的工作中,我们确定了虾肝胰腺中 PirB vp的细胞受体。肝胰腺裂解物的配体印迹分析表明,PirB vp亚基识别两个 60 和 70 kDa 的糖蛋白条带(Gc60 和 Gc70)。通过阴离子交换层析分离肝胰腺裂解物,获得的三个主要级分包含公认的 Gc60 和 Gc70 蛋白条带。LC-MS/MS 表明,β-氨基己糖苷酶亚基 β 和粘蛋白样 5 AC 分别对应于 60 和 70 kDa 的条带,它们似乎在肝胰腺的上皮细胞中表达。用粪肠球菌的O-糖苷酶处理肝胰腺裂解物的糖苷内切酶抑制了 PirB vp的结合。总之,这些结果表明 PirB vp相互作用的相关性 对虾急性肝胰腺坏死病的发病机制与肝胰腺有关。