title: Isolation of inositol 1,4,5-trisphosphate receptor-associating proteins and selective knockdown using RNA interference.
abstract: Inositol 1,4,5-trisphosphate (IP(3)) receptors (IP(3)Rs) are IP(3)-gated Ca(2+) release channels localized on intracellular Ca(2+) stores and play a role in the generation of complex patterns of intracellular Ca(2+) signals. We show herein experimental protocols for the identification of associating proteins of IP(3)R isoforms from various cells and tissues using affinity column chromatography and for the specific knockdown of the expression of IP(3)R isoforms and their associating proteins using RNA interference. These methods will provide clues to understand the exact nature of how the signaling complex contributes to the generation of spatio-temporal patterns of intracellular Ca(2+) signals.
authors.affiliation: Laboratory for Developmental Neurobiology, Brain Science Institute, RIKEN, Saitama, Japan.
authors.forename: Akihiro
authors.initials: A
authors.lastname: Mizutani
authors.forename: Katsuhiro
authors.initials: K
authors.lastname: Kawaai
authors.forename: Chihiro
authors.initials: C
authors.lastname: Hisatsune
authors.forename: Hideaki
authors.initials: H
authors.lastname: Ando
authors.forename: Takayuki
authors.initials: T
authors.lastname: Michikawa
authors.forename: Katsuhiko
authors.initials: K
authors.lastname: Mikoshiba
mesh_terms: D000818:Animals; D002460:Cell Line; D006801:Humans; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011506:Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D034622:RNA Interference
chemical_list: D053496:Inositol 1,4,5-Trisphosphate Receptors; D020033:Protein Isoforms; D011506:Proteins