Identification of formylglycine in sulfatases by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry |
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Authors: | Peng Jianhe Schmidt Bernhard von Figura Kurt Dierks Thomas |
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Affiliation: | Institut für Biochemie und Molekulare Zellbiologie, Abt. Biochemie II, Universit?t G?ttingen, Heinrich-Düker-Weg 12, Germany. jpeng@gwdg.de |
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Abstract: | C(alpha)-Formylglycine, the catalytic amino acid residue in the active site of sulfatases, is generated by post-translational modification of a cysteine or serine residue. We describe a highly sensitive procedure for the detection of C(alpha)-formylglycine-containing peptides in tryptic digests of sulfatase proteins. The protocol is based on the formation of hydrazone derivatives of C(alpha)-formylglycine-containing peptides when using dinitrophenylhydrazine as a matrix for matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOFMS). The hydrazone derivatives desorb and ionize with high efficiency and can be detected in the sub-femtomole range. The presence of C(alpha)-formylglycine is indicated by a mass increment of 180.13 u, corresponding to the hydrazone moiety, and also by a unique C-terminal fragment ion, characteristic of sulfatases, that becomes prominent in MALDI post-source decay mass spectra of the hydrazone derivatives. |
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Keywords: | sulfatase formylglycine protein modification matrix‐assisted laser desorption/ionization time‐of‐flight mass spectrometry dinitrophenylhydrazine |
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