Comprehensive analysis of platelet glycoprotein Ibα ectodomain glycosylation.
Comprehensive analysis of platelet glycoprotein Ibα ectodomain glycosylation.
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DOI：
10.1016/j.jtha.2023.01.009
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发表时间：
2023-04
影响因子：
10.4
通讯作者：
Malaker, Stacy A.
中科院分区：
文献类型：
--
作者：
Hollenhorst, Marie A.;Tiemeyer, Katherine H.;Mahoney, Keira E.;Aoki, Kazuhiro;Ishihara, Mayumi;Lowery, Sarah C.;Rangel-Angarita, Valentina;Bertozzi, Carolyn R.;Malaker, Stacy A.
关键词：
血小板糖蛋白（GP） Ibα是结合血管性血友病因子的GPIb-IX-V复合物的主要配体结合亚基。GPIbα高度糖基化，其聚糖被认为在血小板清除、血管性血友病因子结合和免疫性血小板减少综合征的靶抗原中发挥关键作用。尽管它在血小板生物学中很重要，但GPIbα的糖基化谱尚未得到很好的表征。本研究的目的是综合分析GPIbα氨基酸的糖基化位点（糖位点）和糖基结构。对重组表达的或从人血小板中纯化的GPIbα外结构域进行Western blot、质谱糖组学和质谱糖肽分析，以确定糖位和附着聚糖的结构。我们发现了多种N型和o型聚糖，包括唾液聚糖、Tn抗原、T抗原和ABO(H)血型抗原。在重组蛋白的分析中，我们鉴定出62个独特的o -糖位点。在分析从血小板中纯化的内源性蛋白时，我们鉴定出48个独特的o -糖位点和1个n -糖位点。GPIbα粘蛋白结构域紧密o糖基化。糖位点也位于大糖肽结构域和机械感觉结构域内。这项对GPIbα糖基化的综合分析为进一步研究GPIbα聚糖的功能和结构作用奠定了基础。
Platelet glycoprotein (GP) Ibα is the major ligand-binding subunit of the GPIb-IX-V complex that binds von Willebrand factor. GPIbα is heavily glycosylated, and its glycans have been proposed to play key roles in platelet clearance, von Willebrand factor binding, and as target antigens in immune thrombocytopenia syndromes. Despite its importance in platelet biology, the glycosylation profile of GPIbα is not well characterized. The aim of this study was to comprehensively analyze GPIbα amino acid sites of glycosylation (glycosites) and glycan structures. GPIbα ectodomain that was recombinantly expressed or that was purified from human platelets was analyzed by Western blot, mass spectrometry glycomics, and mass spectrometry glycopeptide analysis to define glycosites and the structures of the attached glycans. We identified a diverse repertoire of N- and O-glycans, including sialoglycans, Tn antigen, T antigen, and ABO(H) blood group antigens. In the analysis of the recombinant protein, we identified 62 unique O-glycosites. In the analysis of the endogenous protein purified from platelets, we identified 48 unique O-glycosites and 1 N-glycosite. The GPIbα mucin domain is densely O-glycosylated. Glycosites are also located within the macroglycopeptide domain and mechanosensory domain. This comprehensive analysis of GPIbα glycosylation lays the foundation for further studies to determine the functional and structural roles of GPIbα glycans.
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DOI：
10.1073/pnas.1511175112
发表时间：
2015-11-24
影响因子：
11.1
作者：
Goth, Christoffer K.;Halim, Adnan;Schjoldager, Katrine T. -B. G.
通讯作者：
Schjoldager, Katrine T. -B. G.
影响因子：
20.3
作者：
Jansen, A. J. Gerard;Josefsson, Emma C.;Hoffmeister, Karin M.
通讯作者：
Hoffmeister, Karin M.
影响因子：
1.5
作者：
Hou, M;Stockelberg, D;Wadenvik, H
通讯作者：
Wadenvik, H
影响因子：
4.1
作者：
JUDSON, PA;ANSTEE, DJ;CLAMP, JR
通讯作者：
CLAMP, JR
影响因子：
20.3
作者：
Curtis, BR;Edwards, JT;Aster, RH
通讯作者：
Aster, RH