Prokaryotic expression of bone sialoprotein and identification of casein kinase II phosphorylation sites.
Prokaryotic expression of bone sialoprotein and identification of casein kinase II phosphorylation sites.
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DOI：
10.1016/j.bbrc.2005.05.124
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发表时间：
2005-07
影响因子：
3.1
通讯作者：
F. Saad;E. Salih;Lívius Wunderlich;R. Flückiger;M. Glimcher
中科院分区：
文献类型：
--
作者：
F. Saad;E. Salih;Lívius Wunderlich;R. Flückiger;M. Glimcher
骨涎蛋白是一种细胞外非胶原性酸性蛋白，在骨矿化和骨重建中发挥作用。它的表达仅限于矿化组织，并受到各种翻译后修饰，包括磷酸化和糖基化。我们在原核系统中表达了牛骨唾液酸蛋白的全长和半结构域，并确定了酪蛋白激酶II的磷酸化位点。N端自动固相测序确定了四个磷酸化多肽：残基28-38(LEDSPEENGVFK)、51-86(FYPELKRFAVQSSSPDSPSPEENGDSPSPEEEEEETSP)、151-165(EDESPDEEEEEEEEEE)和295-305(GRGYDSPYDGQD)。在4个多肽中鉴定出9个磷酸丝氨酸。其中7个在N端(S31、S64、S66、S67、S75、S76和S86)，2个在C端(S154和S300)。
Bone sialoprotein is an extracellular noncollagenous acidic protein that plays a role in bone mineralization and remodeling. Its expression is restricted to mineralized tissues and is subjected to variety of posttranslational modifications including phosphorylation and glycosylation. We have expressed the full-length and half domains of bovine bone sialoprotein in a prokaryotic system and identified the phosphorylation sites of casein kinase II. The N-terminal automated solid-phase sequencing defined four phosphorylated peptides: residues 28–38 (LEDSPEENGVFK), 51–86 (FYPELKRFAVQSSSPDSPSPEENGNGDSPSPEEEEEEEETSP), 151–165 (EDESPDEEEEEEEEEE), and 295–305 (GRGYDSPYDGQD). Nine phosphoserines were identified within the four peptides. Seven of them were in the N-terminus (S31, S64, S66, S67, S75, S76, and S86) and two were in the C-terminus (S154 and S300) of the protein.