Aitken A, Howell S, Jones D, Madrazo J, Martin H, Patel Y, Robinson K
Laboratory of Protein Structure, National Institute for Medical Research, Mill Hill, London, UK.
Mol Cell Biochem. 1995 Aug-Sep;149-150:41-9. doi: 10.1007/BF01076562.
This report compares the ability of individual members of the 14-3-3 protein family to inhibit particular protein kinase C (PKC) isoforms. We also show that two of these 14-3-3 isoforms (alpha and delta) specific to mammalian and avian brain are in vivo post-translationally modified forms of beta and zeta respectively. The presence of this modification enhances the activity of 14-3-3 as an inhibitor of protein kinase C nearly two fold. A method for analysing isoforms of 14-3-3 on acid-urea gels is also described. This permits the complete separation of all major isoforms and their unequivocal identification by a range of isoform specific antisera. The activity of recombinant 14-3-3 and isoforms renatured by a novel method after separation by reverse phase HPLC are compared. The effects of diacylglycerol and the phorbol ester, PMA (phorbol 1 2-myristate 13 acetate) on the inhibition suggest that one of the sites of interaction of 14-3-3 may be the cysteine-rich (C1) domain in PKC.
本报告比较了14-3-3蛋白家族各成员抑制特定蛋白激酶C（PKC）同工型的能力。我们还表明，哺乳动物和鸟类大脑特有的这两种14-3-3同工型（α和δ）分别是β和ζ的体内翻译后修饰形式。这种修饰的存在使14-3-3作为蛋白激酶C抑制剂的活性增强了近两倍。还描述了一种在酸性尿素凝胶上分析14-3-3同工型的方法。这使得所有主要同工型能够完全分离，并通过一系列同工型特异性抗血清进行明确鉴定。比较了重组14-3-3和通过反相高效液相色谱分离后用新方法复性的同工型的活性。二酰基甘油和佛波酯PMA（佛波醇1,2-十四烷酸酯13-乙酸酯）对抑制作用的影响表明，14-3-3的相互作用位点之一可能是PKC中富含半胱氨酸的（C1）结构域。
