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Use of nonspecific cleavage products for protein sequence analysis as shown on calcyclin isolated from human granulocytes

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Journal of the American Society for Mass Spectrometry

Abstract

In this paper, analysis strategies developed for a sequencing problem concerning the identification of an S100 protein isolated from human granulocytes are discussed. The analysis of a trypsinized lyophilized sample suggested the presence of a number of peptides which are non-tryptic in origin. During purification of proteins from cell lysates nonspecific cleavage can be observed which may reflect biological processes and can become an unavoidable analytical problem. Current mass spectrometric software is evaluated for the analysis of nonspecific digests in this context. Matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS), high-performance liquid chromatography (HPLC)-MS/MS, and selected ion monitoring (SIM)-MS/MS have been used for peptide analysis and in addition HPLC-MS was carried out for protein analysis leading to the detection of an N-terminal modification of the protein. The success of the study is mainly due to the careful investigation of nonspecific cleavage products. Data obtained from the routine mass spectrometric analysis of an in-gel-digest allowed the identification of this protein as S100 calcium-binding protein A6—calcyclin whose expression in granulocytes has not been described so far.

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Authors and Affiliations

  1. Institute of Medical Physics and Biophysics, University of Münster, Münster, Germany

    Simone König, Martin Zeller & Jasna Peter-Katalinic

  2. Institute of Experimental Dermatology, University of Münster, Münster, Germany

    Johannes Roth, Clemens Sorg & Thomas Vogl

  3. Integrierte Functionelle Genomik, von-Esmarch-Str. 56, 48149, Münster, Germany

    Simone König

Authors
  1. Simone König
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  2. Martin Zeller
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  3. Jasna Peter-Katalinic
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  4. Johannes Roth
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  5. Clemens Sorg
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  6. Thomas Vogl
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Correspondence to Simone König.

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König, S., Zeller, M., Peter-Katalinic, J. et al. Use of nonspecific cleavage products for protein sequence analysis as shown on calcyclin isolated from human granulocytes. J Am Soc Mass Spectrom 12, 1180–1185 (2001). https://doi.org/10.1016/S1044-0305(01)00300-2

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  • DOI: https://doi.org/10.1016/S1044-0305(01)00300-2

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