Methods in molecular biology (Clifton, N.J.)

Calmodulin-binding proteome in the brain.

PMID 20058172


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.