Toward a general chemical method for rapidly mapping multi-protein complexes.
Ru(II)(bpy2)32+Cl2, ammonium persulfate, and visible light irradiation has been shown to rapidly and efficiently cross-link several interacting proteins. However, this methodology has not yet been used to map the architecture of large multi-protein complexes. In this study, this chemistry is applied to the crystallographically characterized yeast proteasome. The data obtained demonstrate both the method's increased generality and fidelity in comparison to traditional bifunctional cross-linking reagents, while also highlighting the future need for developing better analytical techniques to separate cross-linked products.
Denison
Carilee
C
Center for Biomedical Inventions, Department of Internal Medicine, University of Texas Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390-8573, USA.
Kodadek
Thomas
T
D003432:Cross-Linking Reagents / Q000737:chemistry*; D013058:Mass Spectrometry; D008958:Models, Molecular; D046912:Multiprotein Complexes / Q000032:analysis* / Q000737:chemistry; D046988:Proteasome Endopeptidase Complex / Q000032:analysis* / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000378:metabolism
D003432:Cross-Linking Reagents; D046912:Multiprotein Complexes; D046988:Proteasome Endopeptidase Complex