Wisniewski J P, Monsigny M, Delmotte F M
Laboratoire des Glycoconjugués et des Lectines Endogènes, CNRS, Orléans, France.
Biochimie. 1994;76(2):121-8. doi: 10.1016/0300-9084(94)90003-5.
Lectins associated with the bacterial cell surface of Rhizobium lupini strain LL13 were evidenced by erythrocyte agglutination, by aggregation of neoglycoprotein coated beads and by spectrofluorimetry using fluoresceinylated neoglycoproteins. At pH 5.0, a specific binding of the fluorescein-labelled neoglycoprotein bearing alpha-L-fucose was observed. The binding of this labelled neoglycoprotein is a saturable phenomenon and is inhibited by the same unlabelled neoglycoprotein. Extracts of R lupini obtained by disrupting a bacterial pellet through a French press were stabilized at pH 5.6 by gel filtration and purified to homogeneity by affinity chromatography on Agarose A4 substituted with alpha-L-fucose. A protein with a M(r) approximately 19,000 was specifically eluted from this affinity column with L-fucose. Isoelectric focusing of this sample yielded a single band with pI near 6.7. This protein specifically aggregated L-Fuc-BSA-coated microspheres. The results obtained in the present study indicate that we have purified from Rhizobium lupini strain LL13, a L-fucose binding protein as a lectin.
通过红细胞凝集、新糖蛋白包被珠的聚集以及使用荧光素化新糖蛋白的荧光光谱法，证实了与羽扇豆根瘤菌菌株LL13细菌细胞表面相关的凝集素。在pH 5.0时，观察到携带α-L-岩藻糖的荧光素标记新糖蛋白的特异性结合。这种标记新糖蛋白的结合是一种可饱和现象，并受到相同未标记新糖蛋白的抑制。通过法国压榨机破坏细菌沉淀获得的羽扇豆根瘤菌提取物，通过凝胶过滤在pH 5.6下稳定，并通过用α-L-岩藻糖取代的琼脂糖A4上的亲和色谱法纯化至同质。用L-岩藻糖从该亲和柱上特异性洗脱了一种分子量约为19,000的蛋白质。该样品的等电聚焦产生了一条pI接近6.7 的单带。这种蛋白质特异性聚集了L-Fuc-BSA包被的微球。本研究获得的结果表明，我们已从羽扇豆根瘤菌菌株LL13中纯化出一种作为凝集素的L-岩藻糖结合蛋白。
