title: In vivo protein cross-linking.
abstract: In the cell, homo- and hetero-associations 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 for the selection of 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: Département de Biologie Cellulaire et Infection, Institut Pasteur, Unité de Signalisation et Pathogenèse, 25 rue du Docteur Roux, F-75015, Paris, France, fagou@pasteur.fr.
authors: Fabrice
authors: F
authors: Agou
authors: Michel
authors: M
authors: Véron
mesh_terms: D000595:Amino Acid Sequence; D002463:Cell Membrane Permeability; D006801:Humans; D009701:Nucleoside-Diphosphate Kinase / Q000737:chemistry*; D010455:Peptides / Q000737:chemistry / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*
chemical_list: D010455:Peptides; D011506:Proteins; D009701:Nucleoside-Diphosphate Kinase