摘要
蛋白质磷酸化是生物体内存在的一种普遍的调节方式 ,在细胞信号传递中占有极重要的地位 .质谱已逐渐被人们认为是挑战这一领域的有利工具 .综述了目前利用质谱技术分析磷酸化蛋白质的方法 ,包括利用固定化的金属亲和层析柱、抗体和化学标签技术富集目的分子 ,肽片段质量图和前体离子扫描 ( precusorionscans)等技术检测磷酸化肽段 ,串联质谱对磷酸化肽段测序鉴定磷酸化位点 ,以及引入质量标签对蛋白质的磷酸化水平进行定量等 .虽然现在已经有很多可行的方法用于分析磷酸化蛋白质 。
Protein phosphorylation is a universal regulatory way in organism, and it is a key event in cell signal transduction. Mass spectrometry has emerged as an increasingly viable tool for this task. The methodologies currently available for the analysis of phosphoproteins by mass spectrometry were summarized, including enrichment of compounds of interest using immobilized metal affinity chromatography, antibody and chemical tagging techniques, detection of phosphopeptides using mass mapping and precursor ion sequencing, localization of phosphorylation sites by peptide sequencing, and quantitation of phosphorylation by the introduction of mass tags. Despite the variety of powerful analytical methods that are now available, complete characterization of the phosphorylation state of a protein isolated in small quantities from a biological sample remains far from routine.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2003年第3期350-356,共7页
Progress In Biochemistry and Biophysics
基金
国家重点基础研究发展规划项目 ( 973) ( 0 0 1CB5 10 2 0 4
G19980 5 12 13)
国家自然科学基金资助项目 ( 3990 60 0 )~~
