140 Mouse brain proteins identified by Ca2+-calmodulin affinity chromatography and tandem mass spectrometry

被引:60
作者
Berggård, T
Arrigoni, G
Olsson, O
Fex, M
Linse, S
James, P
机构
[1] Lund Univ, Dept Prot Technol, Wallenberglab, SE-22100 Lund, Sweden
[2] Lund Univ, Dept Biophys Chem, SE-22100 Lund, Sweden
[3] Lund Univ, Dept Cell & Mol Biol, Sect Mol Signaling, SE-22184 Lund, Sweden
关键词
calmodulin; calcium; protein-protein interactions; proteomics; brain;
D O I
10.1021/pr050421l
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Calmodulin is an essential Ca2+-binding protein that binds to a variety of targets that carry out critical signaling functions. We describe the proteomic characterization of mouse brain Ca2+-calmodulin-binding proteins that were purified using calmodulin affinity chromatography. Proteins in the eluates from four different affinity chromatography experiments were identified by 1-DE and in-gel digestion followed by LC-MS/MS. Parallel experiments were performed using two related control-proteins belonging to the EF-hand family. After comparing the results from the different experiments, we were able to exclude a significant number of proteins suspected to bind in a nonspecific manner. A total of 140 putative Cal(2+)-calrnodulin-binding proteins were identified of which 87 proteins contained calmodulin-binding motifs. Among the 87 proteins that contained calmodulin-bincling motifs, 48 proteins have not previously been shown to interact with calmodulin and 39 proteins were known calmodulin-binding proteins. Many proteins with ill-defined functions were identified as well as a number of proteins that at the time of the analysis were described only as ORFs. This study provides a functional framework for studies on these previously uncharacterized proteins.
引用
收藏
页码:669 / 687
页数:19
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