Wang Y H, Fiol C J, DePaoli-Roach A A, Bell A W, Hermodson M A, Roach P J
Department of Biochemistry, Indiana University School of Medicine, Indianapolis 46223.
Anal Biochem. 1988 Nov 1;174(2):537-47. doi: 10.1016/0003-2697(88)90053-x.
A simple procedure is described for determining the location of phosphorylation sites in phosphopeptides. The method employs measurement of 32P-labeled inorganic phosphate release during Edman degradation cycles using a gas-phase sequencer. The procedure is based on extracting peptides and inorganic phosphate from portions of the sample filter at strategic cycles in the sequence analysis followed by determination of the relative amounts of phosphate and phosphopeptide. One advantage of this technique is the very high recovery of the phosphate associated with the peptide, 80-97% in this study. In the course of this work, it was also found that phosphoserine residues themselves caused reduced efficiency of the Edman degradation as compared with unesterified serine residues. The present procedure has the merit of being simple and easy to apply.
本文描述了一种用于确定磷酸肽中磷酸化位点位置的简单方法。该方法利用气相测序仪在埃德曼降解循环过程中测量32P标记的无机磷酸盐释放量。该程序基于在序列分析的关键循环中从样品滤膜的部分中提取肽和无机磷酸盐，然后测定磷酸盐和磷酸肽的相对量。该技术的一个优点是与肽相关的磷酸盐回收率非常高，本研究中为80-97%。在这项工作过程中，还发现与未酯化的丝氨酸残基相比，磷酸丝氨酸残基本身会导致埃德曼降解效率降低。本方法具有简单易行的优点。
