Peñalver M C, O'Connor J E, Martinez J P, Gil M L
Departamento de Microbiología y Ecología, Facultad de Farmacia, Universitat de Valencia, Spain.
Infect Immun. 1996 Apr;64(4):1146-53. doi: 10.1128/iai.64.4.1146-1153.1996.
Aspergillus fumigatus conidia exhibited the ability to bind purified human fibronectin, whereas mycelial forms did not bind the ligand, as detected by an indirect immunofluorescence assay with an antifibronectin polyclonal antibody after incubation of the cells with fibronectin. Flow cytometry confirmed that binding of the ligand to conidia was dose dependent and saturable. Pretreatment of the cells with trypsin markedly reduced binding, which suggested a protein nature for the binding sites present at the surface of conidia. Intact conidia were also able to adhere to fibronectin or antifibronectin antibodies, a significant reduction (from 88 to 92%) in the binding of conidia was noticed, thus suggesting that adhesion to the immobilized ligand was specific. Analysis by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western immunoblotting with fibronectin and antifibronectin antibody of whole conidial homogenates and 2-mercaptoethanol extracts from isolated conidial cell walls allowed identification, among the complex array of protein and glycoprotein species present in both cell-free preparations, of two polypeptides with apparent molecular masses of 23 and 30 kDa which specifically interact with fibronectin.
烟曲霉分生孢子表现出结合纯化人纤连蛋白的能力，而菌丝体形式则不结合该配体，这是在用纤连蛋白孵育细胞后，通过使用抗纤连蛋白多克隆抗体的间接免疫荧光测定法检测到的。流式细胞术证实配体与分生孢子的结合是剂量依赖性且可饱和的。用胰蛋白酶预处理细胞可显著降低结合，这表明分生孢子表面存在的结合位点具有蛋白质性质。完整的分生孢子也能够粘附于纤连蛋白或抗纤连蛋白抗体，观察到分生孢子的结合显著减少（从88%至92%），因此表明对固定化配体的粘附是特异性的。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳以及用纤连蛋白和抗纤连蛋白抗体对整个分生孢子匀浆和从分离的分生孢子细胞壁提取的2-巯基乙醇提取物进行Western免疫印迹分析，在两种无细胞制剂中存在的复杂蛋白质和糖蛋白种类中，鉴定出两种表观分子量分别为23 kDa和30 kDa的多肽，它们与纤连蛋白特异性相互作用。
