Unique properties of prothrombin in the bullhead shark Heterodontus japonicus: the first report of purification and characterization of a blood coagulation factor in Chondrichthyes
Unique properties of prothrombin in the bullhead shark Heterodontus japonicus: the first report of purification and characterization of a blood coagulation factor in Chondrichthyes
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牛头鲨Heterodontus japonicus中凝血酶原的独特特性：软骨鱼中凝血因子的纯化和表征的第一份报告
DOI：
10.1007/s00360-022-01472-3
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Tsutsui Shigeyuki
中科院分区：
文献类型：
--
作者：
Matsui Shintaro;Nakamura Osamu;Tsutsui Shigeyuki
凝血酶原是凝血系统的丝氨酸蛋白酶前体。本研究利用RNA-Seq技术对软骨鱼类牛头鲨（Heterodontus shark）凝血酶原的一级结构进行了分析，并从血浆中纯化了凝血酶原蛋白。发现牛头鲨凝血酶原由四个结构域组成，与已报道的哺乳动物同源物一样。鲨鱼凝血酶原的氨基酸序列中发现了两个被激活的X因子裂解的精氨酸残基，但凝血酶或meizothrobin的两个裂解位点中只有一个是保守的。鲨鱼凝血酶原的SDS-PAGE表观分子量为110 kDa，而其氨基酸序列的表观分子量为65 kDa。在鲨鱼凝血酶原的第79、108、121、179、199、507和527位天冬酰胺残基处发现了潜在的N-糖基化位点，用N-糖苷酶处理使分子量降低至65 kDa。这表明，与仅含有7 kDa N-聚糖的人类凝血酶原相反，鲨鱼的凝血酶原是高度N-糖基化的。本研究首次报道了软骨鱼凝血因子的纯化和表征。
Prothrombin is a serine protease precursor of the blood coagulation system. In this study, the primary structure of prothrombin of a cartilaginous fish, bullhead shark (Heterodontus japonicus), was determined using RNA-Seq and the protein was purified from the blood plasma. Bullhead shark prothrombin was found to be comprised of four domains, as in the case of reported mammalian homologues. Two arginine residues that should be cleaved by activated factor X were found in the amino acid sequence of the shark prothrombin, but only one of the two cleavage sites for thrombin or meizothrombin was conserved. The apparent molecular mass of the shark prothrombin on SDS-PAGE was 110 kDa, whereas that of its amino acid sequence was 65 kDa. Potential N-glycosylation sites were found at 79th, 108th, 121st, 179th, 199th, 507th, and 527th asparagine residues in the shark prothrombin, and treatment with N-glycosidase reduced the molecular mass to 65 kDa. This indicates that, in contrast to human prothrombin, which has only 7-kDa N-glycans, the prothrombin of the shark is highly N-glycosylated. This study is the first to report on the purification and characterization of blood coagulation factors in a cartilaginous fish.
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大鼠凝血酶原的纯化和性质。
DOI：
10.1016/s0021-9258(18)99400-7
发表时间：
1967
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
L. Li;R. Olson
通讯作者：
R. Olson
多个酶位点参与凝血酶原蛋白水解产物调节凝血酶活性的证据。
影响因子：
3.8
作者：
F. Shi;P. Hogg;D. Winzor;C. Jackson
通讯作者：
C. Jackson
牛凝血酶原的纯化和性质。
影响因子：
2.9
作者：
J. Ingwall;H. Scheraga
通讯作者：
H. Scheraga
影响因子：
7.5
作者：
W. Wu;J. Suttie
通讯作者：
W. Wu;J. Suttie
影响因子：
20.3
作者：
Whelihan, Matthew F.;Zachary, Vicentios;Mann, Kenneth G.
通讯作者：
Mann, Kenneth G.