Monitoring conformational changes in protein complexes using chemical cross-linking and Fourier transform ion cyclotron resonance mass spectrometry: the effect of calcium binding on the calmodulin-melittin complex.
Calmodulin is an EF hand calcium binding protein. Its binding affinities to various protein/peptide targets often depend on the conformational changes induced by the binding of calcium. One such target is melittin, which binds tightly to calmodulin in the presence of calcium, and inhibits its function. Chemical cross-linking combined with Fourier transform ion cyclotron resonance mass spectrometry has been employed to investigate the coordination of calmodulin and melittin in the complex at different concentrations of calcium. This methodology can be used to monitor structural changes of proteins induced by ligand binding, and study the effects these changes have on non- covalent interactions between proteins. Cross-linking results indicate that the binding place of the first melittin in the calcium free calmodulin form is the same as in the calcium loaded calmodulin/melittin complex.
Novak
Petr
P
Institute of Microbiology, Academy of Sciences of the Czech Republic, Prague, Czech Republic.
Havlicek
Vladimir
V
Derrick
Peter J
PJ
Beran
Kyle A
KA
Bashir
Sajid
S
Giannakopulos
Anastassios E
AE
D000595:Amino Acid Sequence; D002118:Calcium / Q000378:metabolism*; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D002234:Carbodiimides / Q000737:chemistry; D003432:Cross-Linking Reagents / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D008555:Melitten / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013388:Succinimides / Q000737:chemistry
C518491:1-ethyl-3-(3-(diethylamino)propyl)carbodiimide; D002147:Calmodulin; D002234:Carbodiimides; D003432:Cross-Linking Reagents; D013388:Succinimides; D008555:Melitten; D002118:Calcium; C019358:disuccinimidyl suberate