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Neuroproteomics pp 181–190Cite as

Calmodulin-Binding Proteome in the Brain

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 566))

Summary

Calcium dyshomeostasis is involved in neuropathological conditions such as traumatic brain injury (TBI), stroke, and neurodegenerative diseases. Under such conditions in the brain, calmodulin (CaM), a Ca2+ sensor, mediates critical signaling functions through binding and regulating a diverse population of downstream targets referred to as calmodulin-binding proteins (CaMBPs). We developed a CaM-affinity capture method followed by reversed-phase liquid chromatography tandem mass spectrometry (RPLC-MSMS) to study the calcium-dependent CaM-binding proteome in rat brain. A total of 69 potential CaMBPs were identified by this proteomic technique, of which 26 were known CaMBPs and 43 were putative novel CaMBPs. This study shows that the CaM-affinity capture when coupled with tandem mass spectrometry may serve as an effective tool toward constructing a brain CaM-binding proteomic network. The general methods described here can be applied to study possible alternations of calmodulin-binding proteome in neurological, neurodegenerative, and psychiatric disorders.

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References

  1. Haeseleer, F., Palczewski, K. (2002) Calmodulin and Ca2+-binding proteins (CaBPs): variations on a theme. Adv Exp Med Biol. 514, 303–317.

    Article  PubMed  CAS  Google Scholar 

  2. Bouché, N., Yellin, A., Snedden, W.A., Fromm, H. (2004) Plant-specific calmodulin-binding proteins. Annu Rev Plant Biol. 56, 435–466.

    Article  Google Scholar 

  3. Benaim, G., Villalobo, A. (2002) Phosphorylation of calmodulin. Functional implications. Eur J Biochem. 269, 3619–3631.

    Article  PubMed  CAS  Google Scholar 

  4. Xia, Z., Storm, D.R. (2005) The role of calmodulin as a signal integrator for synaptic plasticity. Nat Rev Neurosci. 6, 267–276.

    Article  PubMed  CAS  Google Scholar 

  5. Xia, Z., Choi, E.J., Wang, F., Blazynski, C., Storm, D.R. (1993) Type I calmodulin-sensitive adenylyl cyclase is neural specific. J Neurochem. 60, 305–311.

    Article  PubMed  CAS  Google Scholar 

  6. O’Day, D.H. (2003) CaMBOT: profiling and characterizing calmodulin-binding proteins. Cell Signal. 15, 347–354.

    Article  PubMed  Google Scholar 

  7. Reddy, V.S., Ali, G.S., Reddy, A.S. (2002) Genes encoding calmodulin-binding proteins in the Arabidopsis genome. J Biol Chem. 277, 9840–9852.

    Article  PubMed  CAS  Google Scholar 

  8. Zhu, H., Bilgin, M., Bangham, R., Hall, D., Casamayor, A., Bertone, P., Lan, N., et al. (2001) Global analysis of protein activities using proteome chips. Science 293, 2101–2105.

    Article  PubMed  CAS  Google Scholar 

  9. Shen, X., Valencia, C.A., Szostak, J.W., Dong, B., Liu, R. (2005) Scanning the human proteome for calmodulin-binding proteins. Proc Natl Acad Sci USA 102, 5969–5974.

    Article  PubMed  CAS  Google Scholar 

  10. Zhang, Z., Ottens, A.K., Golden, E.C., Hayes, R.L., Wang, K.K.W. (2006) Using calmodulin-affinity capture to study the rat brain calmodulin binding proteome and its vulnerability to calpain and caspase proteolysis. Calcium Binding Proteins 2, 125–134.

    Google Scholar 

  11. Ottens, A.K., Kobeissy, F.H., Wolper, R.A., Haskins, W.E., Hayes, R.L., Denslow, N.D., Wang, K.K.W. (2005) A multidimensional differential proteomic platform using dual-phase ion-exchange chromatography – polyacrylamide gel electrophoresis/reversed-phase liquid chromatography tandem mass spectrometry. Anal Chem. 77, 4836–4845.

    Article  PubMed  CAS  Google Scholar 

  12. Tabb, D.L., McDonald, W.H., Yates, J.R. (2002) DTASelect and contrast: tools for assembling and comparing protein identifications from shotgun proteomics. J Proteome Res. 1, 21–26.

    Article  PubMed  CAS  Google Scholar 

  13. Wang, K.K.W., Villalobo, A., Roufogalis, B.D. (1989) Calmodulin-binding proteins as calpain substrates. Biochem J. 262, 693–706.

    PubMed  CAS  Google Scholar 

  14. O’Day, D.H., Myre, M.A. (2004) Calmodulin-binding domains in Alzheimer’s disease proteins: extending the calcium hypothesis. Biochem Biophys Res Commun. 320, 1051–1054.

    Article  PubMed  Google Scholar 

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Acknowledgments

We wish to thank Stephen F. Larner for insightful discussion and editing assistance. This work was supported by the Department of Defense grant DAMMED-03-1-0066, and NIH grants R01 NS049175-01-A1 and R01 NS051431.

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Authors and Affiliations

  1. Center for Neuroproteomics and Biomarkers Research, Department of Psychiatry, McKnight Brain Institute of the University of Florida, Gainesville, FL, USA

    Zhiqun Zhang, Firas H. Kobeissy & Kevin K. W. Wang

  2. Departments of Anatomy & Neurobiology and Biochemistry, Virginia Commonwealth University, Richmond, VA, USA

    Andrew K. Ottens

  3. Department of Psychiatry, McKnight Brain Institute of the University of Florida, Gainesville, FL, USA

    Juan A. Martínez

  4. Center for Innovative Research, Banyan Biomarkers Inc., Alachua, FL, USA

    Kevin K. W. Wang

Authors
  1. Zhiqun Zhang
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  2. Firas H. Kobeissy
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  3. Andrew K. Ottens
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  4. Juan A. Martínez
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  5. Kevin K. W. Wang
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Corresponding author

Correspondence to Zhiqun Zhang .

Editor information

Editors and Affiliations

  1. Dept. Anatomy & Neurobiology, Virginia Commonwealth University School of Medicine, Richmond, U.S.A.

    Andrew K. Ottens

  2. Center of Innovative Research, Banyan Biomarkers, Inc. Center of Innovative Research, Alachua, U.S.A.

    Kevin K.W. Wang

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Zhang, Z., Kobeissy, F.H., Ottens, A.K., Martínez, J.A., Wang, K.K.W. (2009). Calmodulin-Binding Proteome in the Brain. In: Ottens, A., Wang, K. (eds) Neuroproteomics. Methods in Molecular Biology, vol 566. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-562-6_12

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  • DOI: https://doi.org/10.1007/978-1-59745-562-6_12

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-934115-84-8

  • Online ISBN: 978-1-59745-562-6

  • eBook Packages: Springer Protocols

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