Structural analysis of the complex between calmodulin and full-length myelin basic protein, an intrinsically disordered molecule.
Myelin basic protein (MBP) is present between the cytoplasmic leaflets of the compact myelin membrane in both the peripheral and central nervous systems, and characterized to be intrinsically disordered in solution. One of the best-characterized protein ligands for MBP is calmodulin (CaM), a highly acidic calcium sensor. We pulled down MBP from human brain white matter as the major calcium-dependent CaM-binding protein. We then used full-length brain MBP, and a peptide from rodent MBP, to structurally characterize the MBP-CaM complex in solution by small-angle X-ray scattering, NMR spectroscopy, synchrotron radiation circular dichroism spectroscopy, and size exclusion chromatography. We determined 3D structures for the full-length protein-protein complex at different stoichiometries and detect ligand-induced folding of MBP. We also obtained thermodynamic data for the two CaM-binding sites of MBP, indicating that CaM does not collapse upon binding to MBP, and show that CaM and MBP colocalize in myelin sheaths. In addition, we analyzed the post-translational modifications of rat brain MBP, identifying a novel MBP modification, glucosylation. Our results provide a detailed picture of the MBP-CaM interaction, including a 3D model of the complex between full-length proteins.
Department of Biochemistry, University of Oulu, PO Box 3000, 90014, Oulu, Finland.
Viivi
V
Majava
Chaozhan
C
Wang
Matti
M
Myllykoski
Salla M
SM
Kangas
Sung Ung
SU
Kang
Nobuhiro
N
Hayashi
Peter
P
Baumgärtel
Anthony M
AM
Heape
Gert
G
Lubec
Petri
P
Kursula
D000818:Animals; D001921:Brain; D002147:Calmodulin / Q000737:chemistry*; D002417:Cattle; D002478:Cells, Cultured; D006801:Humans; D008024:Ligands; D051379:Mice; D008958:Models, Molecular; D004676:Myelin Basic Protein / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D017510:Protein Folding; D013552:Swine; D013816:Thermodynamics
D002147:Calmodulin; D008024:Ligands; D004676:Myelin Basic Protein