Skip to main content
. Author manuscript; available in PMC: 2015 May 12.
Published in final edited form as: Cell Mol Life Sci. 2009 Feb;66(3):526–536. doi: 10.1007/s00018-008-8362-1

Figure 1.

Figure 1

Collapsin response-mediator protein-2 (CRMP-2) is a calmodulin-binding protein (CaMBP). (A) Identification of CRMP-2 as a putative CaMBP by tandem mass spectrometry (MS/MS). The rat brain CaM-binding proteome was visualized by Coomassie blue staining. Each visible band was excised, in-gel digested and analyzed by RPLC-MS/MS. The position of the CRMP-2 band, matching the peptide sequences, is boxed. The identified CRMP-2 peptides are underlined. (B) Immunoblot of CRMP-2 as a putative CaMBP from rat brain lysate. CaM-agarose flow through and eluate from a rat brain lysate were separated by SDS-PAGE and analyzed by immunoblot for CRMP-2. (C) Immunoblot of CaM agarose elution of recombinant GST-CRMP-2 and GST. The same amount of recombinant GST-CRMP-2 and GST (2 μg) was run through the CaM-agarose column and eluted by EDTA. The eluates were separated by SDS-PAGE and probed with an anti-GST antibody. (D) CaM-overlay experiments with recombinant GST, GST-CRMP-2 and calcineur-in. Equal amounts of recombinant GST-CRMP-2, GSTand bovine brain calcineurin were analyzed by Western blotting, and probed with 20 ng/ml biotinylated bovine CaM, in the presence of 2 mM CaCl2 or 2 mM EDTA.