Skip to main content

Site-Specific Mutagenesis and Protein Engineering Approach to the Molecular Mechanism of Calcium Signal Transduction by Calmodulin

  • Conference paper
Calcium and Calcium Binding Proteins

Part of the book series: Proceedings in Life Sciences ((LIFE SCIENCES))

Abstract

Calmodulin is an ubiquitous, small and acidic protein involved in calcium signal management in all eukaryotic cells (Van Eldik et al. 1982; Cox et al. 1984). The cell, when stimulated, transiently increases its cytosolic calcium concentrations from approximately 0.1 μM to 10 μM. In this range, calmodulin binds calcium ions, modifies its conformation and thereby modifies the function of calmodulin binding proteins. Therefore, calmodulin is a key element in the transduction of a quantitative calcium signal into a cellular response (Klee et al. 1980; Kilhoffer et al. 1983).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Chapter
USD 29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Similar content being viewed by others

References

  • Brodin P, Grundstrom T, Hofmann T, Drakenberg T, Thulen E, Forsen S (1986) Expression of bovine intestinal calcium binding proteins from a synthetic gene in Escherichia coli and characterization of the product. Biochemistry 25:5371–5378

    Article  PubMed  CAS  Google Scholar 

  • Cox JA, Comte M, Stein EA (1982) Activation of human erythrocyte Ca2+-dependent Mg2+-activated ATPase by calmodulin and calcium. Proc Natl Acad Sci USA 79:4265–4269

    Article  PubMed  CAS  Google Scholar 

  • Cox JA, Comte M, Malnoë A, Burger D, Stein EA (1984) Mode of action of the regulatory calmodulin. Met Ions Biol Syst 17:215–293

    CAS  Google Scholar 

  • Craig TA, Watterson DM, Prendergast FP, Haiech J, Roberts DM (1987) Site-specific mutagenesis of the alpha-helices of calmodulin: effects of altering a charge cluster in the helix that links the two halves of calmodulin. J Biol Chem 262:3278–3284

    PubMed  CAS  Google Scholar 

  • Crouch TH, Klee CB (1980) Positive cooperative binding of calcium to bovine brain calmodulin. Biochemistry 19:3692–3698

    Article  PubMed  CAS  Google Scholar 

  • Edelman AM, Takio K, Blumenthal DK, Hansen RS, Walsh KA, Titani K, Krebs EG (1985) Characterization of the calmodulin-binding and catalytic domains in skeletal muscle myosin light chain kinase. J Biol Chem 260:11275–11285

    PubMed  CAS  Google Scholar 

  • Forsen S, Vogel HJ, Drakenberg T (1986) In: Cheung WY (ed) Calcium and cell function. Vol 6. Academic Press, NY, pp 113–157

    Google Scholar 

  • Haiech J, Demaille JG (1981) Supramolecular organization of regulatory proteins into calcisomes: a model of the concerted regulation by calcium ions and cyclic adenosine 3′:5′-monophosphate in eukaryotic cells. In: Holzer (ed) Metabolic interconversion of enzyme. Springer, Berlin Heidelberg New York, pp 303–313 (Proc Life Sci)

    Google Scholar 

  • Haiech J, Sallantin J (1985) Computer search of calcium binding sites in a gene data bank: use of learning techniques to build an expert system. Biochemie 5:555–560

    Article  Google Scholar 

  • Haiech J, Klee CB, Demaille JG (1981) Effects of cations on affinity of calmodulin for calcium: ordered binding of calcium ions allows the specific activation of calmodulin stimulated enzymes. Biochemistry 20:3890–3897

    Article  PubMed  CAS  Google Scholar 

  • Huang CY, Chou V, Chock PB, Wang JH, Sharma RK (1981) Mechanism of activation of cyclic nucleotide phosphodiesterase: requirement of the binding of four Ca2+ to calmodulin for activation. Proc Natl Acad Sci USA 78:871–874

    Article  PubMed  CAS  Google Scholar 

  • Kilhoffer MC, Haiech J, Demaille JG (1983) Ion binding to calmodulin. Mol Cell Biochem 51: 33–54

    Article  PubMed  CAS  Google Scholar 

  • Klee CB, Crouch TH, Richman PG (1980) Calmodulin. Ann Rev Biochem 49:489–515

    Article  PubMed  CAS  Google Scholar 

  • Klotz IM (1985) Ligand-receptor interactions: facts and fantasies. Q Rev Biophysics 18:227–259

    Article  CAS  Google Scholar 

  • Lukas TJ, Iverson DB, Schleicher M, Watterson DM (1984) Structural characterization of a higher plant calmodulin. Plant Physiol 75:788–795

    Article  PubMed  CAS  Google Scholar 

  • Lukas TJ, Marshak DR, Watterson DM (1985a) Drug-protein interactions: isolation and characterization of covalent adducts of phenoxybenzamine and calmodulin. Biochemistry 26:151–157

    Article  Google Scholar 

  • Lukas TJ, Wiggins ME, Watterson DM (1985b) Amino acid sequence of a novel calmodulin from the unicellular alga Chlamydomonas. Plant Physiol 78:477–483

    Article  PubMed  CAS  Google Scholar 

  • Lukas TJ, Burgess WH, Prendergast FG, Lau W, Watterson DM (1986) Calmodulin binding domains: characterization of a phosphorylation and calmodulin binding site from myosin light chain kinase. Biochemistry 25:1458–1464

    Article  PubMed  CAS  Google Scholar 

  • Malnoë A, Cox SA, Stein EA (1982) Ca2+-dependent regulation of calmodulin binding and adenylate cyclase activation in bovine cerebellar membranes. Biochim Biophys Acta 714:84–92

    Article  PubMed  Google Scholar 

  • Marshak DR, Clarke M, Roberts DM, Watterson DM (1984) Structural and functional properties of calmodulin from the eukaryotic microorganism Dictyostelium discoideum. Biochemistry 23: 2891–2899

    Article  PubMed  CAS  Google Scholar 

  • Milos M, Schaer JJ, Comte M, Cox JA (1986) Calcium-proton and calcium-magnesium antagonisms in calmodulin: microcalorimetric and Potentiometrie analyses. Biochemistry 25:6279–6287

    Article  PubMed  CAS  Google Scholar 

  • Roberts DM, Crea R, Malecha M, Alvarado-Urbina G, Chiarello RH, Watterson DM (1985) Chemical synthesis and expression of a calmodulin gene designed for site-specific mutagenesis. Biochemistry 24:5090–5098

    Article  PubMed  CAS  Google Scholar 

  • Schaefer WH, Lukas TJ, Blair IA, Schultz JE, Watterson DM (1987) Amino acid sequence of a novel calmodulin from Paramecium tetraurelia that contains dimethyllysine in the first domain. J Biol Chem 262:1025–1029

    PubMed  CAS  Google Scholar 

  • Van Eldik LJ, Zendegui JG, Marshak DR, Watterson DM (1982) Calcium-binding proteins and the molecular basis of calcium action. Int Rev Cytol 77:1–61

    Article  PubMed  Google Scholar 

  • Walsh M, Stevens JC, Oikowa K, Kay CM (1978) Circular dichroism studies on Ca2+ dependent protein modulator oxidized with N-chlorosuccinimide. Biochemistry 17:3928–3930

    PubMed  CAS  Google Scholar 

  • Wang CL (1985) A note on Ca2+ binding to calmodulin. Biochem Biophys Res Com 130:426–430

    Article  PubMed  CAS  Google Scholar 

  • Watterson DM, Sharief F, Vanaman TC (1980) The complete amino-acid sequence of the Ca2+-dependent modulator protein (calmodulin) of bovine brain. J Biol Chem 255:962–975

    PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1988 Springer-Verlag Berlin Heidelberg

About this paper

Cite this paper

Haiech, J., Watterson, D.M. (1988). Site-Specific Mutagenesis and Protein Engineering Approach to the Molecular Mechanism of Calcium Signal Transduction by Calmodulin. In: Gerday, C., Bolis, L., Gilles, R. (eds) Calcium and Calcium Binding Proteins. Proceedings in Life Sciences. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-73042-9_14

Download citation

  • DOI: https://doi.org/10.1007/978-3-642-73042-9_14

  • Publisher Name: Springer, Berlin, Heidelberg

  • Print ISBN: 978-3-642-73044-3

  • Online ISBN: 978-3-642-73042-9

  • eBook Packages: Springer Book Archive

Publish with us

Policies and ethics