Reid Easton A, Cao Zheng, Wang Yilun, Leite Browning Maria L, Newkirk Robert F, Chaudhuri Guatum, Townsel James G
Department of Anatomy and Physiology, Meharry Medical College, 1005 D.B. Todd Blvd., Nashville, TN 37208, USA.
Life Sci. 2003 Jan 10;72(8):961-76. doi: 10.1016/s0024-3205(02)02343-3.
The protein kinase C (PKC) family of enzymes is broadly distributed and has been implicated in a diverse array of cellular functions. Recent evidence supporting PKC involvement in the regulation of the Limulus choline cotransporter prompted us to clone PKC from a Limulus central nervous system (CNS) cDNA library. An Aplysia californica calcium independent PKC (Apl II) cDNA probe was used to screen the library and 5' RACE SMART PCR was used to obtain the full-length sequence. The resulting cDNA, which included 5' and 3' nontranslation regions, was 4675 bp. Analysis of the encoded peptide sequence using the Swiss-prot database revealed at least 58% identity to PKC epsilon. A commercial polyclonal antibody against PKC epsilon was used in Western blots to positively label a 30 kDa protein from Limulus CNS and the expressed fusion protein of the encoded sequence. These data support the presence of a newly identified PKC-like homolog in Limulus which likely represents a PKC epsilon equivalent.
蛋白激酶C（PKC）家族酶广泛分布，并涉及多种细胞功能。最近有证据支持PKC参与鲎胆碱共转运体的调节，这促使我们从鲎中枢神经系统（CNS）cDNA文库中克隆PKC。使用加州海兔钙非依赖性PKC（Apl II）cDNA探针筛选文库，并使用5' RACE SMART PCR获得全长序列。所得的cDNA包括5'和3'非翻译区，长度为4675 bp。使用瑞士蛋白质数据库对编码的肽序列进行分析，结果显示与PKCε的同一性至少为58%。在蛋白质印迹中使用针对PKCε的商业多克隆抗体对来自鲎CNS的30 kDa蛋白质和编码序列的表达融合蛋白进行阳性标记。这些数据支持在鲎中存在一种新鉴定的PKC样同源物，它可能代表PKCε的等效物。
