PMID: 9164853May 15, 1997Paper

Protein kinase C isoenzymes in airway smooth muscle

The Biochemical Journal
B L WebbM A Giembycz

Abstract

The protein kinase C (PKC) isoenzymes expressed by bovine tracheal smooth muscle (BTSM) were identified at the protein and mRNA levels. Western immunoblot analyses reliably identified PKCalpha, PKCbetaI and PKCbetaII. In some experiments immunoreactive bands corresponding to PKCdelta, PKCepsilon and PKCTheta were also labelled, whereas the gamma, eta and zeta isoforms of PKC were never detected. Reverse transcriptase PCR of RNA extracted from BTSM using oligonucleotide primer pairs designed to recognize unique sequences in the PKC genes for which protein was absent or not reproducibly identified by immunoblotting, amplified cDNA fragments that corresponded to the predicted sizes of PKCdelta, PKCepsilon and PKCzeta, which was confirmed by Southern blotting. Anion-exchange chromatography of the soluble fraction of BTSM following homogenization in Ca2+-free buffer resolved two major peaks of activity. Using epsilon-peptide as the substrate, the first peak of activity was dependent upon Ca2+ and 4beta-PDBu (PDBu=phorbol 12, 13-dibutyrate), and represented a mixture of PKCs alpha, betaI and betaII. In contrast, the second peak of activity, which eluted at much higher ionic strength, also appeared to comprise a combination of convent...Continue Reading

Citations

May 26, 2005·European Journal of Pharmacology·Mark BotermanHerman Meurs
Nov 24, 2004·Journal of Applied Physiology·Chi-Ming Hai, Hak Rim Kim
Mar 11, 2003·Journal of Applied Physiology·A CogoJ G Martin
Jun 19, 2001·American Journal of Physiology. Heart and Circulatory Physiology·H ItohK C Kent
Nov 17, 1998·American Journal of Respiratory and Critical Care Medicine·P J Barnes
Aug 2, 2002·American Journal of Respiratory Cell and Molecular Biology·Kristen PageMarc B Hershenson
Apr 21, 2009·Journal of Smooth Muscle Research = Nihon Heikatsukin Gakkai Kikanshi·Hiroyasu SakaiMiwa Misawa
Oct 1, 2003·Respiratory Physiology & Neurobiology·Limei Zhou, Marc B Hershenson

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