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Calcium-Binding Proteins of the EF-Hand Superfamily pp 127–134Cite as

Coordination to Divalent Cations by Calcium-Binding Proteins

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1929))

Abstract

Fourier-transform infrared spectroscopy (FTIR) is a powerful tool for examining the metal coordination of the side chain COO− groups of Glu and Asp on Ca2+-binding proteins in solution. The behavior of COO− symmetric stretch can be investigated by using protein samples in H2O solution. However, it is difficult to obtain information about the behavior of the COO− antisymmetric stretch in H2O solution, because the COO− antisymmetric stretching band overlaps with the amide II band. Therefore, to obtain reliable infrared spectra in the region of COO− antisymmetric stretch, exchangeable protons in the protein should be completely deuterated by incubating the apoprotein dissolved in D2O under mild heating conditions.

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References

  1. Nara M, Tanokura M (2008) Infrared spectroscopic study of the metal-coordinations structures of calcium-binding proteins. Biochem Biophys Res Comm 369:225–239

    Article  CAS  Google Scholar 

  2. Nara M, Morii H, Tanokura M (2013) Coordination to divalent cations by calcium-binding proteins studied by FTIR spectroscopy. Biochim Biophys Acta Biomembr 1828:2319–2327

    Article  CAS  Google Scholar 

  3. Nara M, Morii H, Tanokura M (2013) Infrared study of synthetic peptide analogues of the calcium-binding site III of troponin C: the role of helix F of an EF-hand motif. Biopolymers 99:42–347

    Article  Google Scholar 

  4. Suzuki N, Imai LF, Kato Y, Nagata K, Ohashi Y, Kuchitsu K, Tanokura M, Sakamoto A, Nara M, Nakano M, Yonezawa N (2013) Coordination structures of Mg2+ and Ca2+ in three types of tobacco calmodulins in solution: Fourier transform infrared spectroscopic studies of side chain COO− Groups. Biopolymers 99:472–483

    Article  CAS  Google Scholar 

  5. Nara M, Yumoto F, Kagi H, Tanokura M (2008) Infrared spectroscopic study of the binding of divalent cations to Akazara scallop troponin C: the effect of the methylene side chain of glutamate residues. Biopolymers 89:595–599

    Article  CAS  Google Scholar 

  6. Yumoto F, Nara M, Kagi H, Iwasaki W, Ojima T, Nishita K, Nagata K, Tanokura M (2001) Coordination structures of Ca2+ and Mg2+ in Akazara scallop troponin C in solution. FTIR spectroscopy of side-chain COO− groups. Eur J Biochem 268:6284–6290

    Article  CAS  Google Scholar 

  7. Deacon GB, Phillips RJ (1980) Relationships between the carbon-oxygen stretching frequencies of carboxylato complexes and the type of carboxylate coordination. Coord Chem Rev 33:227–250

    Article  CAS  Google Scholar 

  8. Nakamoto K (1997) Complexes of alkoxides, alcohols, ethers, ketones, aldehydes, esters and carboxylic acids. In: Nakamoto K (ed) Infrared and Raman spectra of inorganic and coordination compounds part B, 5th edn. Wiley, New York, pp 57–62

    Google Scholar 

  9. Tackett JE (1989) FT-IR characterization of metal acetates in aqueous-solution. Appl Spectrosc 43:483–499

    Article  CAS  Google Scholar 

  10. Nara M, Torii H, Tasumi M (1996) Correlation between the vibrational frequencies of the carboxylate group and the types of its coordination to a metal ion: an ab initio molecular orbital study. J Phys Chem 100:19812–19817

    Article  CAS  Google Scholar 

  11. Tanokura M, Yamada K (1984) Heat capacity and entropy changes of calmodulin induced by calcium binding. J Biochem 95:643–649

    Article  CAS  Google Scholar 

  12. Shaw GS (2001) Synthetic calcium-binding peptides. In: Hans HJ (ed) Methods in molecular biology, vol 173. Humana, Totowa, NJ, pp 175–181

    Google Scholar 

  13. Barth A (2000) The infrared absorption of amino acid side chains. Prog Biophys Mol Biol 74:141–173

    Article  CAS  Google Scholar 

  14. Nara M, Tanokura M, Tasumi M (1994) UV-laser induced changes in the infrared spectrum of calmodulin in the presence of a photosensitive Ca2+ chelator. J Mol Struct 324:107–112

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. Department of Chemistry, College of Liberal Arts and Sciences, Tokyo Medical and Dental University, Chiba, Japan

    Masayuki Nara & Hisayuki Morii

  2. Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, University of Tokyo, Tokyo, Japan

    Masaru Tanokura

Authors
  1. Masayuki Nara
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  2. Hisayuki Morii
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  3. Masaru Tanokura
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Corresponding author

Correspondence to Masayuki Nara .

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Editors and Affiliations

  1. Division of Clinical Chemistry and Biochemistry, Department of Pediatrics, University of Zürich, Zürich, Switzerland

    Claus W. Heizmann

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Nara, M., Morii, H., Tanokura, M. (2019). Coordination to Divalent Cations by Calcium-Binding Proteins. In: Heizmann, C. (eds) Calcium-Binding Proteins of the EF-Hand Superfamily. Methods in Molecular Biology, vol 1929. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-9030-6_9

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  • DOI: https://doi.org/10.1007/978-1-4939-9030-6_9

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-9029-0

  • Online ISBN: 978-1-4939-9030-6

  • eBook Packages: Springer Protocols

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