Photoaffinity labeling combined with mass spectrometric approaches as a tool for structural proteomics.
Protein chemistry, such as crosslinking and photoaffinity labeling, in combination with modern mass spectrometric techniques, can provide information regarding protein-protein interactions beyond that normally obtained from protein identification and characterization studies. While protein crosslinking can make tertiary and quaternary protein structure information available, photoaffinity labeling can be used to obtain structural data about ligand-protein interaction sites, such as oligonucleotide-protein, drug-protein and protein-protein interaction. In this article, we describe mass spectrometry-based photoaffinity labeling methodologies currently used and discuss their current limitations. We also discuss their potential as a common approach to structural proteomics for providing 3D information regarding the binding region, which ultimately will be used for molecular modeling and structure-based drug design.
Robinette
David
D
University of North Carolina at Chapel Hill, UNC-Duke Proteomics Core Facility, Department of Biochemistry and Biophysics, CB7028, Chapel Hill, NC 27599, USA. robinett@med.unc.edu
Neamati
Nouri
N
Tomer
Kenneth B
KB
Borchers
Christoph H
CH
D001386:Azides / Q000737:chemistry; D001577:Benzophenones / Q000737:chemistry; D003978:Diazomethane / Q000031:analogs & derivatives / Q000737:chemistry; D008024:Ligands; D013058:Mass Spectrometry / Q000379:methods*; D008958:Models, Molecular; D019920:Photoaffinity Labels; D025941:Protein Interaction Mapping; D040901:Proteomics / Q000379:methods*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014466:Ultraviolet Rays
D001386:Azides; D001577:Benzophenones; D008024:Ligands; D019920:Photoaffinity Labels; D003978:Diazomethane