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Proteome-wide analysis of protein carboxy termini: C terminomics

Nature Methods volume 7, pages 508–511 (2010)Cite this article

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Abstract

As proteome-wide C-terminal sequence analysis has been largely intractable, we developed a polymer-based enrichment approach to profile protein C-terminal peptides by mass spectrometry and identified hundreds of C-terminal peptides in the Escherichia coli proteome. We isotopically labeled GluC protease–digested and undigested samples and identified GluC substrates and their cleavage sites by quantification of neo–C-terminal peptides. Our method thus enables global annotation of protein C-terminal posttranslational modifications, including proteolytic truncations.

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Figure 1: C-terminomics workflow.
Figure 2: C-TAILS profiling of protease cleavage sites.

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Acknowledgements

O.S. acknowledges support from the Deutsche Forschungsgemeinschaft (grants SCHI 871/1-1 and 871/2-1) and the Michael Smith Foundation for Health Research (MSFHR). O.B. acknowledges support from the Swiss National Science Foundation and Canadian Institutes of Health Research. P.F.H. is supported by Postdoctoral Fellowships of the German Academic Exchange Service (DAAD) and the MSFHR. C.M.O. is supported by a Canada Research Chair in Metalloproteinase Proteomics and Systems Biology. This work was supported by a grant from Canadian Institutes of Health Research and from a program grant in Breast Cancer Metastases from the Canadian Breast Cancer Research Alliance with funds from the Canadian Breast Cancer Foundation and the Cancer Research Society as well as with an Infrastructure grant from MSHFR. We thank W. Chen, S. Perry and S. He for LC-MS/MS measurements. The University of British Columbia Centre for Blood Research Mass Spectrometry Suite is supported by the Canada Foundation for Innovation and the MSFHR.

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  1. Oliver Schilling

    Present address: Present address: University of Freiburg, Institute for Molecular Medicine and Cell Research, Freiburg, Germany.,

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, University of British Columbia, Centre for Blood Research, Vancouver, British Columbia, Canada

    Oliver Schilling, Olivier Barré, Pitter F Huesgen & Christopher M Overall

  2. Department of Oral Biological and Medical Sciences, University of British Columbia, Centre for Blood Research, Vancouver, British Columbia, Canada

    Oliver Schilling, Olivier Barré, Pitter F Huesgen & Christopher M Overall

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  1. Oliver Schilling
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  2. Olivier Barré
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Contributions

O.S. and C.M.O. conceived the approach. All authors contributed to experimental design. O.S. performed all experiments, except O.B. and P.F.H. performed the BSA and β-casein peptide enrichment studies, and O.B. contributed to the proteome experiments. O.S. and C.M.O. wrote the paper, and all authors edited the paper. O.S. and P.F.H. prepared the figures. C.M.O. supervised the project and provided grant support.

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Correspondence to Christopher M Overall.

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The authors declare no competing financial interests.

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Supplementary Figure 1 and Supplementary Tables 1–5 (PDF 3067 kb)

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Schilling, O., Barré, O., Huesgen, P. et al. Proteome-wide analysis of protein carboxy termini: C terminomics. Nat Methods 7, 508–511 (2010). https://doi.org/10.1038/nmeth.1467

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