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Structural analysis of the complex between calmodulin and full-length myelin basic protein, an intrinsically disordered molecule

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An Erratum to this article was published on 04 May 2010

Abstract

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.

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Abbreviations

CaM:

Calmodulin

MBP:

Myelin basic protein

MPE:

Myelin protein extract

SAXS:

Small-angle X-ray scattering

bMBP:

Bovine MBP

pMBP:

Porcine MBP

mMBP:

Mouse MBP

SRCD:

Synchrotron radiation circular dichroism spectroscopy

PTM:

Post-translational modification

3DE:

3-Dimensional electrophoresis

IDP:

Intrinsically disordered protein

SC:

Schwann cell

DRG:

Dorsal root ganglion

MM:

Molecular mass

SPR:

Surface plasmon resonance

ITC:

Isothermal titration calorimetry

PNS:

Peripheral nervous system

CNS:

Central nervous system

RT:

Room temperature

FCS:

Fetal calf serum

SL:

Schmidt–Lanterman incisure

CM:

Compact myelin

∆C p :

Heat capacity

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Acknowledgments

This study was supported by the Academy of Finland, the Sigrid Juselius Foundation, the Finnish MS Foundation, the Finnish Cultural Foundation, and the Department of Biochemistry, University of Oulu. Data collection at MAX-Lab and BESSY (project BESSY-09.1.80843) was supported by the European Community-Research Infrastructure Action under the FP6 “Structuring the European Research Area” Programme, contract RII3-CT-2004-506008 (IA-SFS). We thank the staff of beamline I711 at MAX-Lab and the Biocenter Oulu Proteomics core facility for excellent support. The authors declare no conflicts of interest.

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Authors and Affiliations

  1. Department of Biochemistry, University of Oulu, PO Box 3000, 90014, Oulu, Finland

    Viivi Majava, Chaozhan Wang, Matti Myllykoski & Petri Kursula

  2. Department of Anatomy and Cell Biology, Institute of Biomedicine, University of Oulu, Oulu, Finland

    Salla M. Kangas & Anthony M. Heape

  3. Department of Pediatrics, Medical University of Vienna, Vienna, Austria

    Sung Ung Kang & Gert Lubec

  4. Department of Life Science, Tokyo Institute of Technology, Tokyo, Japan

    Nobuhiro Hayashi

  5. Department of Physical Biochemistry, University of Potsdam, Potsdam, Germany

    Peter Baumgärtel

  6. Helmholtz-Zentrum Berlin, BESSY II, Berlin, Germany

    Peter Baumgärtel

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  1. Viivi Majava
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  2. Chaozhan Wang
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  3. Matti Myllykoski
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  4. Salla M. Kangas
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  5. Sung Ung Kang
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  8. Anthony M. Heape
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  9. Gert Lubec
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  10. Petri Kursula
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Correspondence to Petri Kursula.

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V. Majava and C. Wang contributed equally to this work.

An erratum to this article can be found at http://dx.doi.org/10.1007/s00726-010-0583-6

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Majava, V., Wang, C., Myllykoski, M. et al. Structural analysis of the complex between calmodulin and full-length myelin basic protein, an intrinsically disordered molecule. Amino Acids 39, 59–71 (2010). https://doi.org/10.1007/s00726-009-0364-2

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