Specific inhibition of ectodomain shedding of glycoprotein Ibα by targeting its juxtamembrane shedding cleavage site.
Specific inhibition of ectodomain shedding of glycoprotein Ibα by targeting its juxtamembrane shedding cleavage site.
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DOI：
10.1111/jth.12425
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发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Li R
中科院分区：
文献类型：
--
作者：
Liang X;Russell SR;Estelle S;Jones LH;Cho S;Kahn ML;Berndt MC;Bunting ST;Ware J;Li R
GPIbα的胞外结构域脱落是一种蛋白水解事件，其中金属蛋白酶ADAM 17切割Gly 464-Val 465键并将糖帽蛋白释放到血浆中，被认为是介导储存血小板清除的关键步骤。支持性证据主要来自使用ADAM 17抑制剂的研究。然而，由于ADAM 17的广泛底物特异性，缺乏明确的证据。实现GPIbα脱落的底物特异性抑制。开发直接结合GPIbα脱落切割位点周围序列并通过阻断ADAM 17进入切割位点来抑制GPIbα脱落的单克隆抗体。获得了6株不同亲和力的抗GPIb α单克隆抗体。命名为5G 6的原型克隆及其单体Fab片段特异性结合纯化的GPIb-IX复合物、人血小板和表达人GPIbα的转基因鼠血小板。5G 6在人血小板的组成性和诱导性GPIbα脱落中显示出与广泛使用的脱落抑制剂GM 6001相似的抑制效力。它不识别小鼠GPIbα。它也不抑制其他血小板受体的脱落。最后，5G 6结合显示对血小板活化和聚集没有可检测的影响。5G 6特异性抑制GPIbα脱落，对血小板功能无可检测的影响。通过脱落切割位点的大分子结合的底物特异性脱落抑制的方法可适用于经历胞外域脱落的许多其他跨膜受体。
Ectodomain shedding of GPIbα, a proteolytic event in which metalloprotease ADAM17 cleaves the Gly464-Val465 bond and releases glycocalicin to the plasma, is considered a critical step in mediating clearance of stored platelets. Supporting evidence has largely come from studies using ADAM17 inhibitors. However, the definitive proof is lacking due to the broad substrate specificity of ADAM17. To achieve substrate-specific inhibition of GPIbα shedding. Development of monoclonal antibodies that directly bind the sequence around the GPIbα shedding cleavage site and inhibit GPIbα shedding by blocking ADAM17 access to the cleavage site. Six anti-GPIbα monoclonal antibodies with varying binding affinities were obtained. The prototypic clone, designated 5G6, and its monomeric Fab fragment, bind specifically purified GPIb-IX complex, human platelets, and transgenic murine platelets expressing human GPIbα. 5G6 showed similar inhibitory potency as a widely used shedding inhibitor GM6001 in both constitutive and induced GPIbα shedding in human platelets. It does not recognize mouse GPIbα. Nor does it inhibit shedding of other platelet receptors. Finally, 5G6 binding displays no detectable effect on platelet activation and aggregation. 5G6 specifically inhibits GPIbα shedding with no detectable effect on platelet functions. The method of substrate-specific shedding inhibition by macromolecular binding of the shedding cleavage site can be applicable to many other transmembrane receptors undergoing ectodomain shedding.
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DOI：
10.1073/pnas.78.5.2712
发表时间：
1981-01-01
期刊：
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA-BIOLOGICAL SCIENCES
影响因子：
--
作者：
CLEMETSON, KJ;NAIM, HY;LUSCHER, EF
通讯作者：
LUSCHER, EF
DOI：
10.1073/pnas.95.7.3408
发表时间：
1998-03-31
影响因子：
11.1
作者：
Maskos, K;Fernandez-Catalan, C;Bode, W
通讯作者：
Bode, W
影响因子：
20.1
作者：
Bergmeier, W;Piffath, CL;Wagner, DD
通讯作者：
Wagner, DD
影响因子：
20.3
作者：
Canault, Matthias;Duerschmied, Daniel;Wagner, Denisa D.
通讯作者：
Wagner, Denisa D.
影响因子：
15.9
作者：
Li, Conglei;Piran, Siavash;Ni, Heyu
通讯作者：
Ni, Heyu