Abstract
Reduction of labile disulphide bonds on leukocyte cell surface proteins plays a regulatory role in immune cell activation. Here I describe a method for the fast, efficient, and unbiased purification of cell-surface proteins containing such labile disulphide bonds. Free thiols liberated from the reduction of labile disulphide bonds are labeled with biotin, purified, enriched, and subsequently identified using liquid chromatography coupled to tandem mass spectrometry. Both the proteins containing the labile disulphide bonds and the position of bonds within the protein are revealed, thus providing a valuable addition to the immunology or biochemistry toolkit.
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Acknowledgments
I am grateful to Prof. Neil Barclay and Dr. Carmen Coxon for critical review of the manuscript. This work was supported by the MRC (grant references G0400808 and G9826026).
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Metcalfe, C. (2019). A Proteomics Workflow for the Identification of Labile Disulphide Bonds at the Cell Surface. In: Hogg, P. (eds) Functional Disulphide Bonds. Methods in Molecular Biology, vol 1967. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9187-7_3
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DOI: https://doi.org/10.1007/978-1-4939-9187-7_3
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-4939-9187-7
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