从人、猪和兔血浆以及大鼠血清中分离和鉴定抗凝血酶III。

Isolation and characterization of antithrombin III from human, porcine and rabbit plasma, and rat serum.

Koide T

J Biochem. 1979 Dec;86(6):1841-50. doi: 10.1093/oxfordjournals.jbchem.a132706.

Abstract

Human, porcine, rabbit, and rat antithrombin III have been purified by affinity chromatography using heparin-agarose. The amino acid and carbohydrate compositions, amino-terminal sequences, immunological cross-reactivities, and inhibitions of human thrombin were studied. 2. Human, porcine, rabbit, and rat antithrombin III are single-chain glycoproteins containing hexose, glucosamine, and neuraminic acid. 3. The total carbohydrate contents were 17, 16, 14, and 15% for human, porcine, rabbit, and rat antithrombin III, respectively. 4. Molecular weights estimated from the migration in sodium dodecyl sulfate (SDS)-poly-acrylamide gel electrophoresis were 59,000, 58,000, 63,000, and 63,000 for human, porcine rabbit, and rat antithrombin III, respectively. 5. These four proteins have similar amino acid compositions, although some minor differences were noted. 6. Human, porcine, and rabbit antithrombin III have a histidine residue at the amino-terminus, while rat antithrombin III contains an amino-terminal asparagine residue. 7. The amino-terminal sequences up to the first 17 residues showed high homology among the four proteins. 8. Some immunological cross-reactivity was observed only between human and porcine antithrombin III. 9. The apparent dissociation constants (KI) for the complexes between human thrombin and human, porcine, rabbit, and rat antithrombin III were about 1.2 x 10(-10) M, 9.5 X 10 (-9) M, 1.4 X 10(-7) M, and 2.8 X 10(-9) M, respectively.

摘要

人、猪、兔和大鼠的抗凝血酶III已通过肝素琼脂糖亲和层析法纯化。研究了其氨基酸和碳水化合物组成、氨基末端序列、免疫交叉反应性以及对人凝血酶的抑制作用。2. 人、猪、兔和大鼠的抗凝血酶III是含有己糖、氨基葡萄糖和神经氨酸的单链糖蛋白。3. 人、猪、兔和大鼠抗凝血酶III的总碳水化合物含量分别为17%、16%、14%和15%。4. 根据在十二烷基硫酸钠（SDS）-聚丙烯酰胺凝胶电泳中的迁移率估算，人、猪、兔和大鼠抗凝血酶III的分子量分别为59,000、58,000、63,000和63,000。5. 这四种蛋白质具有相似的氨基酸组成，尽管存在一些细微差异。6. 人、猪和兔的抗凝血酶III在氨基末端有一个组氨酸残基，而大鼠抗凝血酶III含有一个氨基末端天冬酰胺残基。7. 四种蛋白质中前17个残基的氨基末端序列具有高度同源性。8. 仅在人和猪抗凝血酶III之间观察到一些免疫交叉反应性。9. 人凝血酶与人、猪、兔和大鼠抗凝血酶III之间复合物的表观解离常数（KI）分别约为1.2×10⁻¹⁰M、9.5×10⁻⁹M、1.4×10⁻⁷M和2.8×10⁻⁹M。