Pseudomonas aeruginosa lipopolysaccharide binds galectin-3 and other human corneal epithelial proteins
Pseudomonas aeruginosa lipopolysaccharide binds galectin-3 and other human corneal epithelial proteins
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DOI：
10.1128/iai.65.7.2747-2753.1997
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发表时间：
1997-07-01
影响因子：
3.1
通讯作者：
Hazlett, LD
中科院分区：
文献类型：
--
作者：
Gupta, SK;Masinick, S;Hazlett, LD
本研究的目的是测试半乳糖凝集素-3是否存在于人角膜上皮细胞中，以及从铜绿假单胞菌ATCC 19660中提纯的脂多糖是否与该动物凝集素和/或另一种人角膜上皮蛋白(S)结合，并通过器官培养确定脂多糖的哪一组分(内核或外核或脂A)在细菌结合中起重要作用。从铜绿假单胞菌ATCC 19660和商业内毒素(血清型10型)分离纯化的内毒素在聚丙烯酰胺凝胶分析中不同，但与印迹的hcEP结合相似。通过固相分析确定结合是一种受体-配体类型的相互作用，因为它既是特异性的，又是可饱和的。用重叠方法鉴定了HCEP中的几种内毒素结合蛋白。用Galectin-3抗体进行的免疫印迹显示，在新鲜分离的和培养的转化的人角膜上皮中都存在这种蛋白。结合抑制实验表明，针对内毒素外核区的抗体和抗半乳糖凝集素抗体在体外对细菌结合有明显的抑制作用。这些数据进一步证明，内毒素是铜绿假单胞菌的一种重要的粘附素，它与HCEP中的蛋白质受体分子结合，内毒素结合蛋白之一是Galectin-3，分子的外核部分似乎对内毒素与眼睛的结合至关重要。
The aim of this study was to test whether galectin-3 is present in human corneal epithelium and whether lipopolysaccharide (LPS) purified from Pseudomonas aeruginosa ATCC 19660 binds to this animal lectin and/or to another human corneal epithelial protein(s) (HCEP) and to confirm which component of LPS (inner or outer core or lipid A) is important in bacterial binding by using the eye in organ culture. LPS isolated and purified from P. aeruginosa ATCC 19660 and a commercial LPS (serotype 10) differed in polyacrylamide gel analysis but bound similarly to blotted HCEP. Binding was determined to be a receptor-ligand type of interaction by the solid-phase assay, because it was both specific and saturable. Several LPS binding proteins in HCEP were identified by an overlay method. Western blotting with antibody against galectin-3 revealed the presence of this protein in both freshly isolated and cultured transformed human corneal epithelium. Binding inhibition assays showed that antibody specific for the outer core region of LPS and an anti-galectin antibody significantly inhibited bacterial binding in vitro. These data provide further evidence that LPS is an important adhesin of P. aeruginosa, that it binds to protein receptor molecules in HCEP, that one of the LPS binding proteins is galectin-3, and that the outer core portion of the molecule appears to be critical for LPS binding to the eye.