Odin (ANKS1A) is a Src family kinase target in colorectal cancer cells.
Odin (ANKS1A) is a Src family kinase target in colorectal cancer cells.
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DOI：
10.1186/1478-811x-6-7
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发表时间：
2008-10-09
期刊：
影响因子：
--
通讯作者：
Feller SM
中科院分区：
文献类型：
--
作者：
Emaduddin M;Edelmann MJ;Kessler BM;Feller SM
Src家族激酶（SFK）与某些结直肠癌（CRC）的发生有关。SFK的一个成员Lck在正常结肠上皮中检测不到，但在CRC的一个亚组中异常表达。虽然SFK已在成纤维细胞和不同类型的免疫细胞中进行了广泛的研究，但其在许多上皮癌中的物理和功能靶点仍然缺乏特征。检测64个CRC细胞系的Lck表达。然后分析表达高水平Lck并且还含有高基础水平酪氨酸磷酸化（pY）蛋白的SW620 CRC细胞以鉴定新的SFK靶。由于已知SFK的SH2结构域通常在被激酶结构域磷酸化后结合底物，因此将LckSH2与14种其他SH2进行比较，以确定其作为亲和层析试剂的适用性。LckSH 2纯化的pY蛋白的质谱分析随后鉴定了几种容易被称为SFK激酶底物的蛋白质，包括corpine、Tom1L1（SRCASM）、GIT 1、波形蛋白和AFAP 1L2（XB 130）。还检测到先前报道为其他酪氨酸激酶底物的其他蛋白质，包括EGF和PDGF受体靶Odin。进一步分析Odin，发现在SW620细胞中抑制SFK活性后含有显著更少的pY，表明它是CRC细胞中以前未知的SFK靶标。用SH2结构域亲和层析快速鉴定CRC细胞中已知和新的SFK靶是可行的。在上皮癌细胞中阐明新的SFK靶点如Odin，预计将导致对癌细胞信号传导机制的新见解，也可能有助于指示用于监测肿瘤细胞对药物治疗反应的新生物标志物。
Src family kinases (SFK) are implicated in the development of some colorectal cancers (CRC). One SFK member, Lck, is not detectable in normal colonic epithelium, but becomes aberrantly expressed in a subset of CRCs. Although SFK have been extensively studied in fibroblasts and different types of immune cells, their physical and functional targets in many epithelial cancers remain poorly characterised. 64 CRC cell lines were tested for expression of Lck. SW620 CRC cells, which express high levels of Lck and also contain high basal levels of tyrosine phosphorylated (pY) proteins, were then analysed to identify novel SFK targets. Since SH2 domains of SFK are known to often bind substrates after phosphorylation by the kinase domain, the LckSH2 was compared with 14 other SH2s for suitability as affinity chromatography reagent. Mass spectrometric analyses of LckSH2-purified pY proteins subsequently identified several proteins readily known as SFK kinase substrates, including cortactin, Tom1L1 (SRCASM), GIT1, vimentin and AFAP1L2 (XB130). Additional proteins previously reported as substrates of other tyrosine kinase were also detected, including the EGF and PDGF receptor target Odin. Odin was further analysed and found to contain substantially less pY upon inhibition of SFK activity in SW620 cells, indicating that it is a formerly unknown SFK target in CRC cells. Rapid identification of known and novel SFK targets in CRC cells is feasible with SH2 domain affinity chromatography. The elucidation of new SFK targets like Odin in epithelial cancer cells is expected to lead to novel insight into cancer cell signalling mechanisms and may also serve to indicate new biomarkers for monitoring tumor cell responses to drug treatments.