Comprehensive identification of disulfide bonds using non-specific proteinase K digestion and CID-cleavable crosslinking analysis methodology for Orbitrap LC/ESI-MS/MS data.
Disulfide bonds are valuable constraints in protein structure modeling. The Cys-Cys disulfide bond undergoes specific fragmentation under CID and, therefore, can be considered as a CID-cleavable crosslink. We have recently reported on the benefits of using non-specific digestion with proteinase K for inter-peptide crosslink determination. Here, we describe an updated application of our CID-cleavable crosslink analysis software and our crosslinking analysis with non-specific digestion methodology for the robust and comprehensive determination of disulfide bonds in proteins, using Orbitrap LC/ESI-MS/MS data.
University of Victoria - Genome British Columbia Proteomics Centre, University of Victoria, Victoria, British Columbia V8Z 7X8, Canada.
Karl A T
KA
Makepeace
University of Victoria - Genome British Columbia Proteomics Centre, University of Victoria, Victoria, British Columbia V8Z 7X8, Canada.
Jason J
JJ
Serpa
University of Victoria - Genome British Columbia Proteomics Centre, University of Victoria, Victoria, British Columbia V8Z 7X8, Canada.
Evgeniy V
EV
Petrotchenko
University of Victoria - Genome British Columbia Proteomics Centre, University of Victoria, Victoria, British Columbia V8Z 7X8, Canada; Department of Biochemistry & Microbiology, University of Victoria, Victoria, British Columbia V8P 5C2, Canada.
Christoph H
CH
Borchers
D002853:Chromatography, Liquid / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry* / Q000378:metabolism; D003545:Cysteine / Q000032:analysis* / Q000378:metabolism; D004220:Disulfides / Q000032:analysis* / Q000378:metabolism; D019286:Endopeptidase K / Q000032:analysis* / Q000378:metabolism; D006801:Humans; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*; D053719:Tandem Mass Spectrometry / Q000379:methods*
D003432:Cross-Linking Reagents; D004220:Disulfides; D019286:Endopeptidase K; D003545:Cysteine
Crosslinking; Disulfide; Mass spectrometry; Zero-length