Use of the photoaffinity cross-linking agent N-hydroxysuccinimidyl-4-azidosalicylic acid to characterize salivary-glycoprotein-bacterial interactions.
Use of the photoaffinity cross-linking agent N-hydroxysuccinimidyl-4-azidosalicylic acid to characterize salivary-glycoprotein-bacterial interactions.
复制标题
使用光亲和交联剂 N-羟基琥珀酰亚胺基-4-叠氮水杨酸来表征唾液-糖蛋白-细菌相互作用。
DOI：
10.1042/bj2340043
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Al-Hashimi,I
中科院分区：
文献类型：
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作者：
Bergey,EJ;Levine,MJ;Reddy,MS;Bradway,SD;Al-Hashimi,I
本研究利用碘化交联剂N-羟基琥珀酰亚胺基-4-叠氮水杨酸（阿萨）研究了人腮腺唾液中富含脯氨酸的糖蛋白（PRG）与血链球菌G9 B之间的特异性相互作用。血链球菌G9 B与~（125）I-ASA-PRG的结合呈饱和动力学，可逆性，可被未标记的PRG抑制。与其他糖蛋白和β-半乳糖苷酶的抑制研究表明，结合是由细菌粘附素介导的，对N-乙酰神经氨酸，半乳糖和N-乙酰半乳糖胺具有特异性。交联后，125 I-ASA-PRG-粘附素复合物可用SDS提取，并通过Sepharose CL-6 B凝胶过滤与未偶联的125 I-ASA-PRG分离。约1%的125 I-ASA-PRG交联到细菌表面。在5%（w/v）-聚丙烯酰胺凝胶上通过SDS/聚丙烯酰胺凝胶电泳/荧光照相术检查125 I-ASA-PRG-粘附素复合物，发现PRG与两种细菌组分结合。这些发现支持了我们先前的建议，即人唾液糖蛋白可以特异性地与口腔链球菌相互作用，并且这些相互作用发生在糖蛋白的碳水化合物单元和细菌细胞表面的凝集素之间。
The present study has utilized the iodinatable cross-linking agent N-hydroxysuccinimidyl-4-azidosalicylic acid (ASA) to examine the specific interaction between the proline-rich glycoprotein (PRG) of human parotid saliva and Streptococcus sanguis G9B. The binding of 125I-ASA-PRG to Streptococcus sanguis G9B displayed saturation kinetics, reversibility and was inhibited by unlabelled PRG. Inhibition studies with other glycoproteins and saccharides indicated that binding was mediated by a bacterial adhesin with specificity towards N-acetylneuraminic acid, galactose, and N-acetylgalactosamine. After cross-linking, the 125I-ASA-PRG-adhesin complex could be extracted with SDS and separated from uncoupled 125I-ASA-PRG by gel filtration on Sepharose CL-6B. Approx. 1% of the 125I-ASA-PRG was cross-linked to the bacterial surface. Examination of the 125I-ASA-PRG-adhesin complex by SDS/polyacrylamide-gel electrophoresis/fluorography on 5% -(w/v)-polyacrylamide gels revealed that PRG was bound to two bacterial components. These findings support our previous suggestion that human salivary glycoproteins can specifically interact with oral streptococci and that these interactions occur between the glycoprotein's carbohydrate units and lectin(s) on the bacterial cell surface.