Donnelly R, Yang K, Omary M B, Azhar S, Black J L
Department of Pharmacology, University of Sydney, Australia.
Am J Respir Cell Mol Biol. 1995 Sep;13(3):253-6. doi: 10.1165/ajrcmb.13.3.7654381.
Protein kinase C (PKC) has been implicated in the control of airway smooth muscle (ASM) tone, and abnormalities in PKC-dependent signaling may be associated with asthma. PKC exists in different isoforms, but the pattern of their expression in ASM has not been previously reported. Accordingly, the purpose of the present study was to identify which isoforms of PKC are expressed in ASM. Tissue samples of canine ASM were homogenized and subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis, followed by immunoblotting with a range of antipeptide antibodies to PKC-alpha, -beta I, -beta II, -gamma, -delta, -epsilon, -eta, -theta, and -zeta. Positive controls were run in parallel with ASM. Immunoblots revealed a mixture of both calcium-dependent and calcium-independent isozymes: PKC-beta I and PKC-beta II were the only conventional isoforms detected; PKC-delta, PKC-epsilon, and the new muscle-specific isoform, PKC-theta, were all expressed in ASM, but the lung-specific isoform, PKC-eta, was not detected. The calcium- and phospholipid-independent isoform, PKC-zeta, was also present. Thus, expression of a wide variety of both calcium-dependent and calcium-independent isoforms suggests a complex, multifunctional role of PKC in ASM.
蛋白激酶C(PKC)与气道平滑肌(ASM)张力的调控有关,PKC依赖信号转导异常可能与哮喘相关。PKC存在不同的同工型,但其在ASM中的表达模式此前尚未见报道。因此,本研究旨在确定PKC的哪些同工型在ASM中表达。将犬ASM组织样本匀浆,进行十二烷基硫酸钠聚丙烯酰胺凝胶电泳,随后用一系列针对PKC-α、-βI、-βII、-γ、-δ、-ε、-η、-θ和-ζ的抗肽抗体进行免疫印迹。阳性对照与ASM平行进行。免疫印迹显示钙依赖性和钙非依赖性同工酶的混合物:PKC-βI和PKC-βII是仅检测到的传统同工型;PKC-δ、PKC-ε以及新的肌肉特异性同工型PKC-θ均在ASM中表达,但未检测到肺特异性同工型PKC-η。钙和磷脂非依赖性同工型PKC-ζ也存在。因此,多种钙依赖性和钙非依赖性同工型的表达表明PKC在ASM中具有复杂的多功能作用。