Manihar S R, Varma K L, Das H R
CSIR Centre for Biochemicals, Delhi University Campus.
Indian J Biochem Biophys. 1990 Dec;27(6):464-70.
Hemagglutinating activity can be identified in the plasma of different species of murrel fish. This activity may be divided into four types according to their agglutinability towards erythrocytes from different sources. Type I plasma agglutinates human blood group A erythrocytes, type II can agglutinate neuraminidase treated human A B O erythrocytes, type III shows no agglutinating activity towards human erythrocytes, while type IV agglutinates human erythrocytes non-specifically. All of them bind to DEAE-cellulose but elute out by different salt concentrations. Type IV plasma is found to be a combination of three separate hemagglutinins, which are separable by sequential binding to human A B O erythrocytes. Blood group A specific lectin activity is purified from this plasma using formalinised A group erythrocytes. The apparent homogeneity of this purified lectin is established by polyacrylamide gel electrophoresis, isoelectric focusing and immunodiffusion. This agglutinin is antigenically identical with that isolated from type I plasma by affinity chromatography on N-acetyl-D-galactosamine coupled to epoxy-activated cellulose column. Their molecular weights are also found to be identical (Mr 140,000) in polyacrylamide gel electrophoresis, having two identical subunits. Forssman glycolipid (0.03 mM) was found to be the most potent inhibitor of agglutination, although Gal beta 1-3 GalNAc (0.09 mM) is also a good inhibitor. Exhaustive dialysis of the purified lectin (hemagglutinin) against EDTA denatures it irreversibly by dissociating it to its subunit structure. Thus human A group agglutinating activity isolated from type I and type IV plasma are identical.
在不同种类的鳢血浆中可鉴定出血凝活性。根据其对不同来源红细胞的凝集能力，这种活性可分为四种类型。I型血浆能凝集人类A血型红细胞，II型能凝集经神经氨酸酶处理的人类ABO红细胞，III型对人类红细胞无凝集活性，而IV型能非特异性地凝集人类红细胞。它们都能与DEAE -纤维素结合，但通过不同盐浓度洗脱。发现IV型血浆是三种不同血凝素的组合，可通过依次与人类ABO红细胞结合而分离。使用经甲醛处理的A血型红细胞从这种血浆中纯化出A血型特异性凝集素活性。通过聚丙烯酰胺凝胶电泳、等电聚焦和免疫扩散确定了这种纯化凝集素的表观均一性。这种凝集素与通过在偶联到环氧活化纤维素柱上的N -乙酰 - D -半乳糖胺上进行亲和层析从I型血浆中分离出的凝集素在抗原性上相同。在聚丙烯酰胺凝胶电泳中还发现它们的分子量相同（Mr 140,000），有两个相同的亚基。发现福斯曼糖脂（0.03 mM）是最有效的凝集抑制物，尽管β1 - 3半乳糖 - N -乙酰半乳糖胺（0.09 mM）也是一种良好的抑制剂。将纯化的凝集素（血凝素）用EDTA进行彻底透析会使其不可逆地变性，使其解离为亚基结构。因此，从I型和IV型血浆中分离出的人类A血型凝集活性是相同的。
