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Identification of phosphorylation sites of proteins by high performance liquid chromatography-electrospray ionization-quadrupole ion trap mass spectrometry

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Abstract

The phosphorylation sites of two phosphorylated proteins, bovine β-casein and myelin basic protein (MBP), were identified by high performance liquid chromatography-electrospray ionization-quadrupole ion trap mass spectrometry (HPLC-ESI-QITMS). The tryptic digest of each protein was separated by HPLC, the molecular weight of each peptide was determined by ESI-QITMS on line, and MS/MS spectrum of each peptide was simultaneously obtained by the combination of collision-induced desorption (CID) technique and tandem mass spectrometry (MS/MS) of QITMS. The phosphorylated peptide was identified by looking into whether the difference between the observed and predicted molecular weights of a peptide is 80 u or its integral multiple. Then the phosphorylation site was identified through manual interpretation of the MS/MS spectrum of the phosphorylated peptide or automatic SEQUEST data base-searching.

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Authors and Affiliations

  1. Shanghai Institute of Biochemistry, Chinese Academy of Sciences, 200031, Shanghai, China

    Fayun Che, Xiaoxia Shao & Qichang Xia

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  1. Fayun Che
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  2. Xiaoxia Shao
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  3. Qichang Xia
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Correspondence to Qichang Xia.

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Che, F., Shao, X. & Xia, Q. Identification of phosphorylation sites of proteins by high performance liquid chromatography-electrospray ionization-quadrupole ion trap mass spectrometry. Sci. China Ser. C.-Life Sci. 43, 561–568 (2000). https://doi.org/10.1007/BF02882276

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  • DOI: https://doi.org/10.1007/BF02882276

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