Prostasin and hepatocyte growth factor B in factor VIIa generation: Serine protease knockdowns in zebrafish.
Prostasin and hepatocyte growth factor B in factor VIIa generation: Serine protease knockdowns in zebrafish.
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VIIA因子产生的前列腺素和肝细胞生长因子B：斑马鱼中的丝氨酸蛋白酶敲低。
DOI：
10.1002/rth2.12428
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发表时间：
2020-10
影响因子：
4.6
通讯作者：
Jagadeeswaran P
中科院分区：
文献类型：
--
作者：
Khandekar G;Iyer N;Jagadeeswaran P
人体内的血液凝结是由组织因子与血浆中激活的凝血因子VII(FVIIa)结合而引起的。以前的研究报道，肝素和激活因子VII(FVII)的蛋白酶负责产生FVIIa。我们的目标是以斑马鱼为模型，确定其他可能激活FVII的蛋白酶。我们用背负式基因敲除的方法筛选了179个编码丝氨酸蛋白酶结构域的基因，以确定斑马鱼FviI激活的相关基因。延长动态凝血酶原时间(KPT)检测基因敲除效应。在初筛中，有21个基因显示KPT延长。在二次筛选中，21个基因中有14个基因呈阳性结果。在第三次筛查中，所有14个基因都显示KPT延长。这14个基因再次被删除，以估计斑马鱼FVIIa的相对水平。6个基因，包括已知基因f10和新的前列腺素和肝细胞生长因子B(Hgfb)，显示出较低的FVIIa水平。Fvii水平只受到f7基因敲除的影响，而不受其他五个基因的敲除的影响。前列腺素和hGFb参与FVIIa的生成。我们推测前列腺素通过直接或间接地发挥丝氨酸蛋白酶活性来激活FVII。由于Hgfb有一个突变的丝氨酸蛋白酶结构域，它可能不会切割Fvii，但可能会与Fvii结合以诱导自身激活。这里开发的方法可以推广到其他大型击落式屏幕的设计。
Blood clotting in humans is initiated by the binding of tissue factor to activated coagulation factor VII (FVIIa) in the plasma. Previous studies have reported that hepsin and factor VII (FVII)‐activating protease are responsible for generating FVIIa. We aimed to identify other proteases that may activate FVII using zebrafish as a model. We screened 179 genes encoding serine protease domains using the piggyback knockdown method to identify genes involved in the activation of zebrafish Fvii. A prolonged kinetic prothrombin time (kPT) assay was used to detect gene knockdown effects. In the primary screen, 21 genes showed prolonged kPT. In the secondary screen, 14 of 21 genes showed positive results. In the tertiary screen, all 14 genes showed prolonged kPT. These 14 genes were knocked down again to estimate relative levels of zebrafish Fviia. Six genes, including known genes, such as f10 and novel prostasin and hepatocyte growth factor B (hgfb), showed lower Fviia levels. Fvii levels were affected only by the knockdown of f7 and not by the knockdown of the other five genes. Prostasin and hgfb are involved in generating Fviia. We hypothesize that prostasin exerts serine protease activity directly or indirectly to activate Fvii. As Hgfb has a mutated serine protease domain, it may not cleave Fvii but may bind to Fvii to induce autoactivation. The approach developed here may be extended to design other large‐scale knockdown screens.
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影响因子：
2.9
作者：
PEDERSEN, AH;LUNDHANSEN, T;PETERSEN, LC
通讯作者：
PETERSEN, LC
影响因子：
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