GONOCOCCAL OPACITY - LECTIN-LIKE INTERACTIONS BETWEEN OPA PROTEINS AND LIPOOLIGOSACCHARIDE
GONOCOCCAL OPACITY - LECTIN-LIKE INTERACTIONS BETWEEN OPA PROTEINS AND LIPOOLIGOSACCHARIDE
复制标题
DOI：
10.1128/iai.63.4.1434-1439.1995
复制
发表时间：
1995-04-01
影响因子：
3.1
通讯作者：
MANDRELL, RE
中科院分区：
文献类型：
--
作者：
BLAKE, MS;BLAKE, CM;MANDRELL, RE
来自我们实验室的先前证据表明，显示不透明菌落表型的淋球菌外膜之间的紧密细胞间粘附发生，因为在一个淋球菌上表达的Opa蛋白粘附到相对细菌的脂寡糖（LOS）（M. S. Blake，p. 51-66，in G. G.杰克逊和H.托马斯编，细菌感染的发病机制，1985年，和M。S. Blake和E. C. Gotschlich，p. 377-400，in M. Inouye，编，Bacterial Outer Membranes as Model Systems，1986），对以前用于确定主要肝去唾液酸糖蛋白受体识别的碳水化合物结构的非竞争性抑制测定进行了修改，以确定结合Opa蛋白的淋球菌LOS结构（R. T.李，目标诊断，Ther. 4：65-84，1991）。在这些测定中使用的LOS碳水化合物是从淋病奈瑟氏菌1291的绿脓菌素LOS突变体纯化的LOS结构，其由K. C. Dudas和M. A. Apicella（Infect. Immun. 56：499-504，1988），并进一步由C. M. John et al.（J.Biol.Chem.266：19303-19311，1991），将纯化的淋球菌Opa蛋白与亲本和突变体LOS中的每一种一起温育，并通过使用Opa特异性单克隆抗体4 B12的直接酶联免疫吸附测定来测量Opa蛋白的结合量。通过测量非竞争性抑制LOS特异性单克隆抗体50%结合的Opa蛋白浓度，间接确定Opa蛋白对每个LOS的亲和力。该浓度与抑制剂的亲和力成反比（R. T.李，有针对性的诊断。4：65-84，1991）。我们的数据表明，测试的淋球菌Opa蛋白对淋球菌新内酯系列LOS上存在的Gal β 1-4GlcNAc残基具有最高亲和力。该亲和力与所报道的主要肝去唾液酸糖蛋白受体与含有末端半乳糖和N-乙酰半乳糖胺的糖缀合物的结合亲和力相当（R. T.李，有针对性的诊断。4：65-84，1991）。后唾液酸化的lactoneoseries LOS，大概在末端半乳糖残基，与Opa蛋白质的相互作用被消融。因此，淋球菌Opa-LOS和哺乳动物上皮细胞脱唾液酸糖蛋白受体-碳水化合物相互作用具有非常相似的特异性。
Previous evidence from our laboratory suggested that the tight intercellular adhesions between the outer membranes of gonococci displaying the opacity colony phenotype occurred because Opa proteins expressed on one gonococcus adhered to the lipooligosaccharide (LOS) of the opposing bacterium (M. S. Blake, p. 51-66, in G. G. Jackson and H. Thomas, ed., The Pathogenesis of Bacterial Infections, 1985, and M. S. Blake and E. C. Gotschlich, p. 377-400, in M. Inouye, ed., Bacterial Outer Membranes as Model Systems, 1986), A noncompetitive inhibition assay used previously to determine the carbohydrate structures recognized by the major hepatic asialoglycoprotein receptor was modified to determine the gonococcal LOS structures that bind Opa proteins (R. T. Lee, Targeted Diagn., Ther. Ser. 4:65-84, 1991). The LOS carbohydrates used in these assays were LOS structures purified from pyocin LOS mutants of Neisseria gonorrhoeae 1291 described by K. C. Dudas and M. A. Apicella (Infect. Immun. 56:499-504, 1988) and further characterized by C. M. John ct al. (J. Biol. Chem. 266:19303-19311, 1991), Purified gonococcal Opa proteins were incubated with each of the parent and mutant LOS, and the amount of binding of Opa proteins was measured by a direct enzyme-linked immunosorbent assay using the Opa-specific monoclonal antibody 4B12. The affinities of the Opa proteins for each of the LOS were determined indirectly by measuring the concentrations of Opa proteins that noncompetitively inhibited 50% of the binding of LOS-specific monoclonal antibodies. This concentration is inversely proportional to the affinity of the inhibitor (R. T. Lee, Targeted Diagn. Ther. Ser. 4:65-84, 1991). Our data suggest that the gonococcal Opa proteins tested had the highest affinity for the Gal beta 1-4GlcNAc residue present on the gonococcal lactoneoseries LOS. This affinity was comparable to that reported for the binding of the major hepatic asialoglycoprotein receptor to glycoconjugates containing terminal galactose and N-acetylgalactosamine (R. T. Lee, Targeted Diagn. Ther. Ser. 4:65-84, 1991). After sialylation of the lactoneoseries LOS, presumably on the terminal galactose residue, the interaction with the Opa proteins was ablated. Therefore, the gonococcal Opa-LOS and mammalian epithelial cell asialoglycoprotein receptor-carbohydrate interactions have quite similar specificities.