Novel platelet-agglutinating protein from a thrombotic thrombocytopenic purpura plasma.
Novel platelet-agglutinating protein from a thrombotic thrombocytopenic purpura plasma.
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来自血栓性血小板减少性紫癜血浆的新型血小板凝集蛋白。
DOI：
10.1172/jci112107
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发表时间：
1985
期刊：
影响因子：
--
通讯作者：
E. Lian
中科院分区：
文献类型：
--
作者：
F. Siddiqui;E. Lian
从血栓性血小板减少性紫癜（TTP）患者的血浆中，经硫酸铵分级分离、DEAE-Sephacel和伴刀豆球蛋白A-Sepharose层析纯化了约2,000倍的新的血小板凝集蛋白（PAP）。在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中，在有和没有还原的情况下，该制备物显示分子量为37，000的主要蛋白条带和分子量为32，000 - 34，000的次要条带。从凝胶洗脱后，只有对应于主带的37，000-mol wt蛋白质诱导血小板凝集。当对来自同一TTP患者的四个正常血浆和恢复血浆进行类似的纯化步骤时，37，000-mol wt主带不存在。125 I-PAP与血小板的结合呈浓度依赖性。在抗凝血酶III、苯甲磺酰氟、腺苷三磷酸双磷酸酶、阿司匹林或前列腺素I2存在下，水蛭素、肝素不抑制PAP诱导的血小板凝集。正常人IgG和TTP患者恢复后IgG均能抑制该酶的活性。在兔中制备的抗37，000-mol wt PAP抗血清对高度纯化的PAP形成单一沉淀素线。在Western免疫印迹中使用该抗血清，在三种TTP血浆中发现37，000-mol wt蛋白条带，其中血小板凝集活性被抗37，000-mol wt PAP IgG抑制。另外两名TTP患者、两名正常受试者、两名特发性血小板减少性紫癜患者和两名弥散性血管内凝血患者的血浆中不存在37，000-mol wt免疫组织化学蛋白条带。这些结果表明，37，000-mol wt PAP仅存在于某些TTP病例中，并且可能是微循环中血小板血栓形成的原因。
A novel platelet-agglutinating protein (PAP) was purified approximately 2,000-fold from the plasma of a patient with thrombotic thrombocytopenic purpura (TTP) by ammonium sulfate fractionation, DEAE-Sephacel and concanavalin A-Sepharose chromatographies. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis, with and without reduction, this preparation revealed a major protein band with a molecular weight of 37,000, and a minor band with a molecular weight of 32,000-34,000. After elution from the gel, only the 37,000-mol wt protein corresponding to the major band induced the platelet agglutination. When four normal plasmas and the recovery plasma from the same TTP patient were subjected to the similar purification steps, the 37,000-mol wt major band was absent. The 125I-PAP bound to the platelets in a concentration-dependent manner. The platelet agglutination induced by PAP was not inhibited by hirudin, heparin in the presence of antithrombin III, phenylmethylsulfonyl fluoride, apyrase, aspirin, or prostaglandin I2. However, it was inhibited by IgG from normal adults and from the same TTP patient after recovery. The anti-37,000-mol wt PAP antiserum prepared in the rabbit formed a single precipitin line against the highly purified PAP. Using this antiserum in the Western immunoblotting, the 37,000-mol wt protein band was found in the three TTP plasmas, of which the platelet-agglutinating activity was inhibited by the anti-37,000-mol wt PAP IgG. The 37,000-mol wt immunoprecipitin band was absent in the plasmas obtained from another two TTP patients, two normal subjects, two patients with idiopathic thrombocytopenic purpura, and two patients with disseminated intravascular coagulation. These results suggest that the 37,000-mol wt PAP is present only in certain cases of TTP, and is likely to be responsible for the formation of platelet thrombi in the microcirculation.
血栓性血小板减少性紫癜患者血浆对血小板和培养的血管内皮细胞的病理影响。
影响因子：
20.3
作者：
Burns,ER;Zucker-Franklin,D
通讯作者：
Zucker-Franklin,D