Calmodulin-binding proteome in the brain.
Calcium dyshomeostasis is involved in neuropathological conditions such as traumatic brain injury (TBI), stroke, and neurodegenerative diseases. Under such conditions in the brain, calmodulin (CaM), a Ca(2+) sensor, mediates critical signaling functions through binding and regulating a diverse population of downstream targets referred to as calmodulin-binding proteins (CaMBPs). We developed a CaM-affinity capture method followed by reversed-phase liquid chromatography tandem mass spectrometry (RPLC-MSMS) to study the calcium-dependent CaM-binding proteome in rat brain. A total of 69 potential CaMBPs were identified by this proteomic technique, of which 26 were known CaMBPs and 43 were putative novel CaMBPs. This study shows that the CaM-affinity capture when coupled with tandem mass spectrometry may serve as an effective tool toward constructing a brain CaM-binding proteomic network. The general methods described here can be applied to study possible alternations of calmodulin-binding proteome in neurological, neurodegenerative, and psychiatric disorders.
Zhang
Zhiqun
Z
Department of Psychiatry, University of Florida, Gainesville, FL, USA. zqzhang@banyanbio.com
Kobeissy
Firas H
FH
Ottens
Andrew K
AK
Martínez
Juan A
JA
Wang
Kevin K W
KK
D000818:Animals; D001923:Brain Chemistry; D002147:Calmodulin / Q000378:metabolism; D002148:Calmodulin-Binding Proteins / Q000032:analysis*; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D006801:Humans; D008297:Male; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley; D053719:Tandem Mass Spectrometry / Q000379:methods
D002147:Calmodulin; D002148:Calmodulin-Binding Proteins; D020543:Proteome