Abstract
Mass spectrometric structural analysis of crosslinked peptides is a powerful method to elucidate the spatial arrangement of polypeptides in protein complexes. Our aim is to develop bifunctional crosslinkers that, after crosslinking protein complexes followed by proteolytic digestion, give rise to crosslinked peptides that can be readily tracked down by mass spectrometry. To this end we synthesized the crosslinker N-benzyliminodiacetoyloxysuccinimid (BID), which yields stable benzyl cation marker ions upon low-energy collisioninduced dissociation (CID) tandem mass spectrometry. Sensitive detection of the marker ion upon low-energy CID is demonstrated with different BID-crosslinked peptide preparations. With BID it becomes possible to retrieve crosslinked and crosslinker-adducted peptides, without the necessity of purifying crosslinked peptides prior to identification. The basic design of this crosslinker can be varied upon, in order to meet specific crosslinking needs.
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Shevchenko, A.; Wilm, M.; Vorm, O.; Mann, M. Anal. Chem. 1996, 68, 850–858.
Last, A. M.; Robinson, C. V. Curr. Opin. Chem. Biol. 1999, 3, 564–570.
Benjamin, D. R.; Robinson, C. V.; Hendrick, J. P.; Hartl, F. U.; Dobson, C. M. Proc. Natl. Acad. Sci. USA 1998, 95, 7391–7395.
Rostom, A. A.; Robinson, C. V. Curr. Opin. Struct. Biol. 1999, 9, 135–141.
Rappsilber, J.; Siniossoglou, S.; Hurt, E. C.; Mann, M. Anal. Chem. 2000, 72, 267–275.
Wallon, G.; Rappsilber, J.; Mann, M.; Serrano, L. Embo. J. 2000, 19, 213–222.
Webb, Y.; Zhou, X.; Ngo, L.; Cornish, V.; Stahl, J.; Erdjumentbromage, H.; Tempst, P.; Rifkind, R. A.; Marks, P. A.; Breslow, R.; Richon, V. M. J. Biol. Chem. 1999, 274, 14280–14287.
Alley, S. C.; Jones, A. D.; Soumillion, P.; Benkovic, S. J. J. Biol. Chem. 1999, 274, 24485–24489.
Luo, Y.; Leszyk, J.; Qian, Y.; Gergely, J.; Tao, T. Biochemistry 1999, 38, 6678–6688.
Wong, S. S. Chemistry of Protein Conjugation and Cross-Linking; CRC Press: Boca Raton FL, 1991.
Chen, X.; Eswaran, D.; Smith, M. A.; Perry, G.; Anderson, V. E. Bioconjug. Chem. 1999, 10, 112–118.
Chen, X.; Chen, Y. H.; Anderson, V. E. Anal. Biochem. 1999, 273, 192–203.
Anderson, G. W.; Zimmermam, J. E.; Callahan, F. M. J. Am. Chem. Soc. 1964, 86, 1839.
Ji, T. H. Meth. Enzymol. 1983, 91, 580–609.
Hunter, E. P.; Lias, S. G. In NIST Chemistry WebBook, NIST Standard Reference Database Number 69; Mallard, W. G., Linstrom, P. J., Eds.; National Institute of Standards and Technology: Gaithersburg MD, 2000; Vol. 69.
Cerny, R. L.; Gross, M. L. In Mass Spectrometry of Peptides; Desiderio, D. M., Ed.; CRC Press: Boca Raton, FL, 1991; pp 289–314.
Tsaprailis, G.; Nair, H.; Somogyi, A.; Wysocki, V. H.; Zhong, W. Q.; Futrell, J. H.; Summerfield, S. G.; Gaskell, S. J. J. Am. Chem. Soc. 1999, 121, 5142–5154.
Tsaprailis, G.; Somogyi, A.; Nikolaev, E. N.; Wysocki, V. H. Int. J. Mass Spectrom 2000, 196, 467–479.
Yalcin, T.; Csizmadia, I. G.; Peterson, M. R.; Harrison, A. G. J. Am. Soc. Mass Spectrom. 1996, 7, 233.
Ambihapathy, K.; Yalcin, T.; Leung, H. W.; Harrison, A. G. J. Mass Spectrom. 1997, 32, 209–215.
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Back, J.W., Hartog, A.F., Dekker, H.L. et al. A new crosslinker for mass spectrometric analysis of the quaternary structure of protein complexes. J Am Soc Mass Spectrom 12, 222–227 (2001). https://doi.org/10.1016/S1044-0305(00)00212-9
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DOI: https://doi.org/10.1016/S1044-0305(00)00212-9