Chances and pitfalls of chemical cross-linking with amine-reactive N-hydroxysuccinimide esters.
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.
Kalkhof
Stefan
S
Department of Pharmaceutical Chemistry and Bioanalytics, Institute of Pharmacy, Martin Luther University Halle-Wittenberg, Wolfgang-Langenbeck-Strasse 4, 06120 Halle (Saale), Germany.
Sinz
Andrea
A
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000737:chemistry*; D003432:Cross-Linking Reagents / Q000737:chemistry*; D004952:Esters / Q000737:chemistry; D007797:Laminin / Q000737:chemistry; D008239:Lysine / Q000737:chemistry; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis / Q000737:chemistry*; D012694:Serine / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods; D013388:Succinimides / Q000737:chemistry*; D013912:Threonine / Q000737:chemistry; D014443:Tyrosine / Q000737:chemistry
D000596:Amino Acids; D003432:Cross-Linking Reagents; D004952:Esters; C478221:LAMB1 protein, human; D007797:Laminin; D010455:Peptides; D013388:Succinimides; C089141:laminin gamma 1; D013912:Threonine; D014443:Tyrosine; D012694:Serine; D008239:Lysine; C001426:N-hydroxysuccinimide