• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biochemistry >Mass Spectrometric Identification of Phosphorylation Sites in Guanylyl Cyclase A and B
【24h】

Mass Spectrometric Identification of Phosphorylation Sites in Guanylyl Cyclase A and B

机译：质谱鉴定鸟苷酸环化酶A和B中的磷酸化位点

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Guanylyl cyclase A and B (GC-A and GC-B) are transmembrane guanylyl cyclase receptors thatnmediate the physiologic effects of natriuretic peptides. Some sites of phosphorylation are known for ratGC-Anand GC-B, but no phosphorylation site information is available for the human homologues. Here, we usednmass spectrometry to identify phosphorylation sites in GC-A and GC-B from both species. Tryptic digests ofnreceptors purified from HEK293 cells were separated and analyzed by nLC-MS-MS. Seven sites ofnphosphorylation were identified in rat GC-A (S497, T500, S502, S506, S510, T513, and S487), and all ofnthese sites except S510 and T513 were observed in human GC-A. Six phosphorylation sites were identified innratGC-B (S513, T516, S518, S523, S526, and T529), and all six sites were also identified in humanGC-B. Fivensites are identical betweenGC-AandGC-B. S487 inGC-Aand T529 inGC-B are novel, uncharacterized sites.nSubstitution of alanine for S487 did not affect initial ligand-dependent GC-A activity, but a glutamatensubstitution reduced activity 20%. Similar levels of ANP-dependent desensitization were observed for thenwild-type, S487A, and S487E forms of GC-A. Substitution of glutamate or alanine for T529 increased orndecreased ligand-dependent cyclase activity of GC-B, respectively, and T529E increased cyclase activity in anGC-B mutant containing glutamates for all five previously identified sites as well. In conclusion, we identifiednand characterized new phosphorylation sites in GC-A and GC-B and provide the first evidence ofnphosphorylation sites within human guanylyl cyclases.
机译：鸟苷酸环化酶A和B（GC-A和GC-B）是介导利钠肽的生理作用的跨膜鸟苷酸环化酶受体。 ratGC-Anand GC-B的某些磷酸化位点是已知的，但尚无人类同源物的磷酸化位点信息。在这里，我们使用nmass光谱法来鉴定两种物种的GC-A和GC-B中的磷酸化位点。分离自HEK293细胞纯化的受体的胰蛋白酶消化物，并通过nLC-MS-MS分析。在大鼠GC-A中鉴定出七个磷酸化位点（S497，T500，S502，S506，S510，T513和S487），并且在人GC-A中观察到了除S510和T513以外的所有这些位点。在inratGC-B中鉴定出六个磷酸化位点（S513，T516，S518，S523，S526和T529），并且在人GC-B中也鉴定了所有六个位点。 GC-A和GC-B之间的Fivensite相同。 S487 inGC-A和T529 inGC-B是新颖的未鉴定位点。n丙氨酸取代S487不会影响初始的依赖配体的GC-A活性，但是谷氨酰胺取代会使活性降低20％。对于野生型，S487A和S487E形式的GC-A，观察到相似水平的ANP依赖性脱敏。谷氨酸或丙氨酸替代T529分别增加了GC-B的配体依赖性环化酶活性，或降低了T529E在所有先前鉴定的五个位点的含有谷氨酸的GC-B突变体中的环化酶活性。总之，我们鉴定并鉴定了GC-A和GC-B中新的磷酸化位点，并提供了人类鸟苷酸环化酶内n磷酸化位点的第一个证据。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号