Tryptic phosphopeptides of 32P-labeled IGFBP-5 from T47D cells separated by HPLC and sequenced by tandem MS.a, tandem MS spectrum of the triply charged.

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binding sites 58 of the 473 unambiguously assigned phosphorylation sites are predicted by Scansite to be sites for binding. 50 of these correspond.

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Presentation transcript:

Tryptic phosphopeptides of 32P-labeled IGFBP-5 from T47D cells separated by HPLC and sequenced by tandem MS.a, tandem MS spectrum of the triply charged parent ion at m/z 983.1 from fraction 21. Tryptic phosphopeptides of 32P-labeled IGFBP-5 from T47D cells separated by HPLC and sequenced by tandem MS.a, tandem MS spectrum of the triply charged parent ion at m/z 983.1 from fraction 21. The sequence matched IGFBP-5-(92–115) where Ser96 is phosphorylated and Met107 is oxidized (shown underlined). The parent ion has been truncated, and the m/z range from 990 to 2,000 has been multiplied by a factor of 30 to show the fragment ions more clearly. Neutral loss of methane sulfenic acid (CH3SOH, −64 Da) is shown. b, tandem MS spectrum of the triply charged parent ion at m/z 886.4 from fraction 31. The sequence matched IGFBP-5-(231–252) where Ser248 is phosphorylated. The deduced presence of oxidized Met234 is shown underlined. Also the Cys243 alkylated with acrylamide (see “Experimental Procedures”) is shown underlined. The m/z range from 900 to 1,400 has been multiplied by a factor of 10 to show the fragment ions more clearly. Mark E. Graham et al. Mol Cell Proteomics 2007;6:1392-1405 © 2007 The American Society for Biochemistry and Molecular Biology