从人血小板中溶解出的血栓素A2/前列腺素H2拮抗剂结合位点的流体动力学特性

Hydrodynamic properties of a thromboxane A2/prostaglandin H2 antagonist binding site solubilized from human platelets.

作者信息

Burch R M, Mais D E, Pepkowitz S H, Halushka P V

出版信息

Biochem Biophys Res Commun. 1985 Nov 15;132(3):961-8. doi: 10.1016/0006-291x(85)91901-1.

DOI:10.1016/0006-291x(85)91901-1

Abstract

The hydrodynamic properties of a binding site for the thromboxane A2/prostaglandin H2 receptor antagonist 9,11-dimethylmethano-11, 12-methano-16-(3-iodo-4-hydroxyphenyl)-13, 14-dihydro-13-aza-15 alpha beta-omega-tetranor-thromboxane A2 (I-PTA-OH) were determined in solubilized membrane proteins from human platelets using the detergent 3-[(3-cholamidopropyl)-dimethylammonio] 1-propane-sulfonate (CHAPS). Gel filtration revealed a Stokes radius of 5.25 +/- 0.37 nm (n=9). Molecular weight determined by gel filtration assuming a spherical protein was 180,000-220,000 Daltons. Sedimentation through sucrose or glycerol gradients revealed a sedimentation coefficient of 6.3 +/- 0.2 Svedberg units (n=5). The molecular weight calculated using the Stokes radius and sedimentation coefficient was 140,000 Daltons. The frictional ratio f/fo was 1.4, corresponding to an axial ratio of 7:1.

摘要

使用去污剂3-[(3-胆酰胺丙基)-二甲基铵基]-1-丙烷磺酸盐（CHAPS），在人血小板的溶解膜蛋白中测定了血栓素A2/前列腺素H2受体拮抗剂9,11-二甲基甲烷-11,12-甲烷-16-(3-碘-4-羟基苯基)-13,14-二氢-13-氮杂-15αβ-ω-四降-血栓素A2（I-PTA-OH）结合位点的流体动力学性质。凝胶过滤显示斯托克斯半径为5.25±0.37nm（n=9）。假设蛋白质为球形，通过凝胶过滤测定的分子量为180,000-220,000道尔顿。通过蔗糖或甘油梯度沉降显示沉降系数为6.3±0.2斯维德伯格单位（n=5）。使用斯托克斯半径和沉降系数计算的分子量为140,000道尔顿。摩擦比f/fo为1.4，对应轴比为7:1。