Mazumder T, Gaur N, Surolia A
Eur J Biochem. 1981 Jan;113(3):463-70. doi: 10.1111/j.1432-1033.1981.tb05086.x.
A lectin has been isolated from Momordica charantia seeds and purified to homogeneity by affinity adsorption on a cross-linked arabinogalactan column and subsequent elution with 0.1 M lactose. In sedimentation velocity experiments an S20,W(O) degrees value of 6.5 S was obtained for the native lectin and 2.7 S for its subunits obtained by dissociation with 0.1% sodium dodecyl sulphate in the presence of 0.1% 2-mercaptoethanol. This lectin has a molecular weight of 120 000, is tetrameric in nature with two pairs of non-identical subunits of molecular weights 28 000 and 30 000. It is a glycoprotein containing 10% hexoses (2% glucose and 8% mannose) and 2.5% hexosamine, all glucosamine. It has two binding sites per molecule and has Ka values of 4.1 X 10(4) M-1 and 1.8 X 10(4) M-1 at 5 degrees C and 25 degrees C respectively for p-nitrophenyl beta-D-galactopyranoside with delta H degrees and delta S degrees values of --28.3 kJ mol-1 and --12.3J mol-1 K-1 respectively. Hemagglutinating activity of M. charantia lectin is inhibited by galactose and other carbohydrates containing the galactopyranosyl residue. From the chemical modification studies tryptophan and tyrosine residues are found to be important for the carbohydrate binding activity of M. charantia lectin.
已从苦瓜种子中分离出一种凝集素,并通过在交联阿拉伯半乳聚糖柱上进行亲和吸附,随后用0.1M乳糖洗脱,将其纯化至同质。在沉降速度实验中,天然凝集素的S20,W(O)°值为6.5S,在0.1%2-巯基乙醇存在下用0.1%十二烷基硫酸钠解离得到的亚基的S20,W(O)°值为2.7S。这种凝集素的分子量为120000,本质上是四聚体,由两对分子量分别为28000和30000的不同亚基组成。它是一种糖蛋白,含有10%的己糖(2%葡萄糖和8%甘露糖)和2.5%的己糖胺,均为葡糖胺。每个分子有两个结合位点,在5℃和25℃下,对对硝基苯基β-D-吡喃半乳糖苷的Ka值分别为4.1×10(4)M-1和1.8×10(4)M-1,ΔH°和ΔS°值分别为-28.3kJmol-1和-12.3Jmol-1K-1。苦瓜凝集素的血凝活性受到半乳糖和其他含有吡喃半乳糖基残基的碳水化合物的抑制。从化学修饰研究中发现,色氨酸和酪氨酸残基对苦瓜凝集素的碳水化合物结合活性很重要。