Drago G A, Colyer J
Department of Biochemistry and Molecular Biology, University of Leeds, United Kingdom.
J Biol Chem. 1994 Oct 7;269(40):25073-7.
A pair of polyclonal antibodies have been produced in rabbits which recognize the two phosphorylated forms of the cardiac muscle protein, phospholamban. The two sites of phosphorylation of this protein are situated on neighboring residues and yet one antibody, PS-16, recognizes the Ser16 phosphoprotein, while the other, PT-17, recognizes the Thr17 phosphoprotein. Neither antibody recognizes phospholamban phosphorylated at "the other site," nor do they recognize free phosphoamino acids or dephosphorylated protein. This represents the most demanding test of the technique of phosphorylation site-specific antibody production to date, which these antibodies have satisfied without ambiguity. These antibodies remain specific for phospholamban in the company of other muscle phosphoproteins and will be invaluable in determining the role of each phosphorylation site in the biology of the heart. They also demonstrate the absolute specificity of phosphorylation site-specific antibodies and augur well for the use of this approach in the study of other phosphoproteins.
已在兔子体内产生了一对多克隆抗体，它们可识别心肌蛋白受磷蛋白（phospholamban）的两种磷酸化形式。该蛋白的两个磷酸化位点位于相邻残基上，但一种抗体PS - 16识别Ser16磷酸化蛋白，而另一种抗体PT - 17识别Thr17磷酸化蛋白。两种抗体均不识别在“另一个位点”磷酸化的受磷蛋白，也不识别游离的磷酸氨基酸或去磷酸化蛋白。这是迄今为止对磷酸化位点特异性抗体生产技术要求最严苛的测试，而这些抗体已毫无歧义地通过了测试。在其他肌肉磷酸化蛋白存在的情况下，这些抗体对受磷蛋白仍具有特异性，对于确定每个磷酸化位点在心脏生物学中的作用将非常宝贵。它们还证明了磷酸化位点特异性抗体的绝对特异性，并为这种方法在其他磷酸化蛋白研究中的应用带来了良好的前景。
