title: Investigation of calmodulin-Peptide interactions using matrix-assisted laser desorption/ionization mass spectrometry.
abstract: In this report, matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) was used to study the binding interactions between calmodulin and two target peptides (melittin and substance P). Various matrix conditions were tested and the less acidic matrix DHAP and THAP were found to favor the survival of the intact calcium-calmodulin as well as the calmodulin-peptide complexes. However, the application of direct MALDI-MS to detect the intact complexes turned out to be very difficult due to the dissociation of the complexes and the formation of nonspecific aggregates. In contrast, the specific binding of the target peptides to calmodulin could be easily deduced using intensity-fading (IF) MALDI-MS. Compared with the nonbinding control, clear reduction in the ion abundances of the target peptides was observed with the addition of calmodulin. Relative binding affinities of different peptides towards the protein could also be estimated using IF-MALDI-MS. This study may extend the application of IF-MALDI-MS in the analysis of noncovalent complexes and offer a perspective into the utility of MALDI-MS as an alternative approach to study the peptides binding to calmodulin.
authors.affiliation: Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun Center of Mass Spectrometry, Changchun, PR China.
authors.forename: Zhaofu
authors.initials: Z
authors.lastname: Wang
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Yu
authors.forename: Meng
authors.initials: M
authors.lastname: Cui
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Liu
authors.forename: Fengrui
authors.initials: F
authors.lastname: Song
authors.forename: Shuying
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D001516:Bees; D002118:Calcium / Q000737:chemistry; D002147:Calmodulin / Q000737:chemistry*; D002417:Cattle; D008555:Melitten / Q000737:chemistry*; D011188:Potassium / Q000737:chemistry; D011485:Protein Binding; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013373:Substance P / Q000737:chemistry*
chemical_list: D002147:Calmodulin; D008555:Melitten; D013373:Substance P; D011188:Potassium; D002118:Calcium