Family with sequence similarity member 20C is the primary but not the only kinase for the small-integrin-binding ligand N-linked glycoproteins in bone.
Family with sequence similarity member 20C is the primary but not the only kinase for the small-integrin-binding ligand N-linked glycoproteins in bone.
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具有序列相似性的家族成员 20C 是骨中小整合素结合配体 N 连接糖蛋白的主要但不是唯一的激酶。
DOI：
10.1096/fj.15-273607
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发表时间：
2016
期刊：
影响因子：
--
通讯作者：
Wang,Xiaofang
中科院分区：
文献类型：
--
作者：
Yang,Xiudong;Yan,Wenjuan;Tian,Ye;Ma,Pan;Opperman,LynneA;Wang,Xiaofang
最近的研究发现，具有序列相似性成员20C的家族(FAM20C)是一种磷酸化小整合素结合配体N-连锁糖蛋白(Siblings)中Ser-X-Glu/Phylo-Ser(PSer)基序中的Ser的激酶。没有活体证据证实这一发现，目前还不清楚FAM20C是否是兄弟姐妹的唯一激酶。我们从Fam20C基因敲除(KO)小鼠中提取骨非胶原蛋白(NCPs)，并对其磷酸化水平进行分析。经阴离子交换层析分离得到富含骨桥蛋白、骨涎蛋白和牙本质基质蛋白-1的组份，并用SDS-PAGE、天然PAGE和免疫印迹分析对其进行分析。KO小鼠NCP的磷酸化水平低于野生型(WT)。在天然凝胶上，来自KO小鼠的兄弟姐妹的迁移率(MR)低于来自WT的兄弟姐妹。小牛肠道磷酸酶处理使兄弟姐妹从WT小鼠转移到与KO相邻的水平，但未能转移后者，这表明KO小鼠的兄弟姐妹失去了磷酸化。质谱仪在KO小鼠的兄弟姐妹中发现了较少的PSERS[包括富含酸性丝氨酸和天冬氨酸的基序(ASARM)多肽的区域]。一个有趣的发现是，KO小鼠Ser-X-Glu基序中的几个PSERS保持了它们的磷酸化，而非Ser-X-Glu基序中的其他几个PSERS则不能。兄弟姐妹中的磷酸化酪氨酸和磷酸化THR似乎与FAM20C无关。我们的结果表明，FAM20C是这两个兄弟姐妹的主要但不是唯一的激酶。-Yang，X.，yan，W.，Tian，Y.，Ma，P.，Opperman，L.A.，Wang，X.家族中序列相似的成员20C是骨骼中小整合素结合配体N-连接糖蛋白的主要但不是唯一的激酶。
Recent studies have identified family with sequence similarity member 20C (FAM20C) as a kinase that phosphorylates the Ser in Ser-X-Glu/phospho-Ser (pSer) motifs in the small-integrin–binding ligand N-linked glycoproteins (SIBLINGs). There is no in vivo evidence that validates this finding, and it is unclear whether FAM20C is the only kinase for SIBLINGs. We extracted bone noncollagenous proteins (NCPs) from Fam20C-knockout (KO) mice and analyzed the phosphorylation levels. The total NCPs were separated into osteopontin-, bone sialoprotein-, and dentin matrix protein-1–enriched fractions by anion-exchange chromatography and analyzed by SDS-PAGE, native PAGE, and Western immunoblot analysis. The NCP phosphorylation level in the KO mice was lower than that in the wild-type (WT). On the native gel, the SIBLINGs from KO mice showed a lower migration rate (Mr) than those from the WT. Calf intestine phosphatase treatment shifted SIBLINGs from the WT mice to the level adjacent to the KO, but failed to shift the latter, suggesting a phosphorylation loss of SIBLINGs in the KO mice. Mass spectrometry identified less pSers in the SIBLINGs from the KO mice [including the region of the acidic Ser- and aspartate-rich motif (ASARM) peptides]. In an intriguing finding, several pSers in the Ser-X-Glu motifs in the KO mice maintained their phosphorylation, whereas several others in non–Ser-X-Glu motifs did not. Phospho-Tyrs and phospho-Thrs in the SIBLINGs did not appear to be associated with FAM20C. Our results indicate that FAM20C is the primary, but not the only, kinase for the SIBLINGs.—Yang, X., Yan, W., Tian, Y., Ma, P., Opperman, L. A., Wang, X. Family with sequence similarity member 20C is the primary but not the only kinase for the small-integrin–binding ligand N-linked glycoproteins in bone.
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