Monitoring of S100 homodimerization and heterodimeric interactions by the yeast two-hybrid system.
The S100 family consists of 19 members, which function as transducers of calcium signals in a tissue-specific manner. Upon calcium binding, the conformation of many S100 proteins changes dramatically. Several hydrophobic residues are exposed, allowing the S100 proteins to interact with their target proteins, and thereby to transduce calcium signals into specific biological responses. To further elucidate the exact contribution of the S100 calciproteins in the calcium signalling pathways, several groups have applied the yeast two-hybrid technology to identify putative target proteins for the various S100 calciproteins. Two-hybrid large screens using S100 proteins as baits have confirmed the biochemical and structural feature of S100, which enable them to form homodimers and the ability of some members to form specific heterodimers in vivo. Yeast two-hybrid investigations have allowed the identification of conserved hydrophobic residues and domains that are crucial for the stabilization of S100 homo- and heterodimers. Furthermore, this method clearly underlines that the homo- and heterodimerization mechanisms differ among the members of the S100 family. However, several lines of evidence strongly suggest that two-hybrid methodology is limited to the analysis of interactions that are calcium-independent, since no target proteins other than S100 family members themselves have been detected with this methodology.
Deloulme
Jean Christophe
JC
Département Réponse et Dynamique Cellulaires du CEA, INSERM, EMI 0104, CEA-Grenoble, 38054 Grenoble Cedex 9, France. jcdeloulme@cea.fr
Gentil
Benoît Jean
BJ
Baudier
Jacques
J
D000595:Amino Acid Sequence; D000818:Animals; D002135:Calcium-Binding Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019281:Dimerization; D020832:EF Hand Motifs; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009418:S100 Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D020798:Two-Hybrid System Techniques
D002135:Calcium-Binding Proteins; D009418:S100 Proteins