Molecular cloning and identification of a serine/threonine protein kinase of the second-messenger subfamily.

来自 NCBI

喜欢 0

阅读量：

248

作者：

P F Jones，T Jakubowicz，F J Pitossi，F Maurer，B A Hemmings

展开

摘要：

A partial cDNA was isolated that encoded a protein kinase, termed rac (related to the A and C kinases). This cDNA was subsequently used to screen libraries derived from the human cell lines MCF-7 and WI38 and led to the isolation of full-length cDNA clones. DNA sequence analysis identified an open reading frame of 1440 base pairs encoding a protein of 480 amino acids (Mr, 55,716). This result was supported by the synthesis of a Mr58,000 protein in an in vitro translation system that used RNA transcribed from cloned cDNAs with SP6 RNA polymerase. The predicted protein contains consensus sequences characteristic of a protein kinase catalytic domain and shows 73% and 68% similarity to protein kinase C and the cAMP-dependent protein kinase, respectively. Northern (RNA) analysis revealed a single mRNA transcript of 3.2 kilobases that varied up to 300-fold between different cell lines. Specific antisera directed towards the carboxyl terminal of the rac protein kinase were prepared and used to identify a protein of Mr59,000 by immunoblotting. A specific protein kinase activity was identified that phosphorylated several substrates in immunoprecipitates prepared with the rac-specific antisera.

展开

关键词：

Biochemistry Protein Phosphorylation Signal Transduction Homology to Protein Kinase A and Protein Kinase C MCF-7 and WI38 Cells

DOI：

10.1073/pnas.88.10.4171

被引量：

1708

年份：

1991