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Chances and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters

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Abstract

In this report we summarize our experiences with the reaction products of N-hydroxysuccinimide (NHS) esters, which are widely used for chemical cross-linking of lysine residues in proteins. We describe the products, which should be scrutinized during data analysis using customized software when NHS esters are employed for chemical cross-linking. Reaction products of NHS esters were observed not only with lysines, but also with serines, tyrosines, and threonines. This report is intended to be a practical guide for those working in the field of chemical cross-linking and mass spectrometry.

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Acknowledgements

This work was supported by grant SI 867/7–1 from the Deutsche Forschungsgemeinschaft (DFG); S.K. was supported by the IPP at the University of Leipzig and the GRK 1026 at the Martin Luther University Halle-Wittenberg. The authors are indebted to Mats Paulsson, Neil Smyth, and Sebastian Haehn for expression and purification of N-terminal laminin ß1 and laminin γ1 chain constructs. Tibor Kohajda is acknowledged for programming the software tool ISOFIND.

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  1. Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Strasse 4, 06120, Halle (Saale), Germany

    Stefan Kalkhof & Andrea Sinz

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  1. Stefan Kalkhof
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  2. Andrea Sinz
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Correspondence to Andrea Sinz.

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Kalkhof, S., Sinz, A. Chances and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters. Anal Bioanal Chem 392, 305–312 (2008). https://doi.org/10.1007/s00216-008-2231-5

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  • DOI: https://doi.org/10.1007/s00216-008-2231-5

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