UNIT 19.16 Determination of Protein Contacts by Chemical Cross-Linking With EDC and Mass Spectrometry
Published Online: 1 FEB 2007
Copyright © 2007 by John Wiley & Sons, Inc.
Lab Protocol Title
Current Protocols in Protein Science
How to Cite
Marekov, L. N. 2007. Determination of Protein Contacts by Chemical Cross-Linking With EDC and Mass Spectrometry. Current Protocols in Protein Science. 47:19.16:19.16.1–19.16.8.
- Published Online: 1 FEB 2007
- Published Print: FEB 2007
This unit describes a method for analyzing protein complexes by chemically cross-linking closely positioned amino acids. The cross-linked products are isolated by SDS-PAGE and the bands of interest are excised. Proteins in the excised gel piece are digested with trypsin and the resulting peptides recovered. Chemically bonded peptides are distinct from the multitude of unmodified single peptides by the presence of two carboxy-terminal ends. The protocol also describes incorporation of 18O at the carboxy-terminal ends and purification and preparation of the peptide mixture for mass spectroscopy.
- protein contacts;
- isotopic exchange;
- mass spectrometry