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Marker null for mass spectrometry of phosphorylation protein and phosphorylation position analysis

机译:磷酸化蛋白质谱分析和磷酸化位置分析的无效标记

摘要

Disclosed is a method of analyzing mass of the phosphoproteins or phosphopeptides and of analyzing phosphorylated positions at a phosphoprotein or phosphopeptide, comprising the steps of: 1) dephosphorylating at least one Ser and/or Thr residue of the phosphoprotein or phosphopeptide; 2) tagging the dephosphorylated amino acid residues with a tag having a R-L-G moiety wherein R is a nucleophilic functional group that selectively bind with dephosphorylated amino acid residues, G is selected from the group consisting of guanidine moiety or protected guanidine moiety such as a mono-N-protected guanidino group, a di-N,N'-protected guanidino group and an N'-protected guanidino group, and L is a linker linking the R and the G; and 3) subjecting the tagged proteins or peptides to mass spectrometry. The method is capable of precisely analyzing mass of phosphoproteins of trace amounts as well as positions of phosphoryated amino acids.
机译:公开了一种分析磷蛋白或磷肽的质量并分析磷蛋白或磷肽上的磷酸化位置的方法,该方法包括以下步骤:1)使磷蛋白或磷肽的至少一个Ser和/或Thr残基去磷酸化; 2)用具有RLG部分的标签标记去磷酸化的氨基酸残基,其中R是与去磷酸化氨基酸残基选择性结合的亲核官能团,G选自胍基部分或受保护的胍基部分,例如单- N-保护的胍基,二-N,N'-保护的胍基和N'-保护的胍基,L是连接R和G的接头。 3)对标记的蛋白质或肽进行质谱分析。该方法能够精确分析痕量磷蛋白的质量以及磷酸化氨基酸的位置。

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