title: In vivo protein cross-linking.
abstract: In the cell, homo- and heteroassociations of polypeptide chains evolve and take place within subcellular compartments that are crowded with many other cellular macromolecules. In vivo chemical cross-linking of proteins is a powerful method to examine changes in protein oligomerization and protein-protein interactions upon cellular events such as signal transduction. This chapter is intended to provide a guide to the selection of the cell-membrane-permeable cross-linkers, the optimization of in vivo cross-linking conditions, and the identification of specific cross-links in a cellular context where the frequency of random collisions is high. By combining the chemoselectivity of the homo-bifunctional cross-linker and the length of its spacer arm with knowledge on the protein structure, we show that selective cross-links can be introduced specifically on either the dimer or the hexamer form of the same polypeptide in vitro as well as in vivo, using the human type B nucleoside diphosphate kinase as a protein model.
authors.affiliation: Departement de Biologie Structurale et Chimie, Institut Pasteur, Paris, France.
authors.forename: Fabrice
authors.initials: F
authors.lastname: Agou
authors.forename: Fei
authors.initials: F
authors.lastname: Ye
authors.forename: Michel
authors.initials: M
authors.lastname: Véron
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry*; D006367:HeLa Cells; D006801:Humans; D008958:Models, Molecular; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry / Q000235:genetics; D011506:Proteins / Q000737:chemistry*
chemical_list: D003432:Cross-Linking Reagents; D011506:Proteins; D009701:Nucleoside-Diphosphate Kinase