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Coordination to Divalent Cations by Calcium-Binding Proteins

  • Masayuki NaraEmail author
  • Hisayuki Morii
  • Masaru Tanokura
Protocol
Part of the Methods in Molecular Biology book series (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.

Key words

FTIR Coordination structure Carboxylate group Synthetic peptide analogue HD exchange 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Masayuki Nara
    • 1
    Email author
  • Hisayuki Morii
    • 1
  • Masaru Tanokura
    • 2
  1. 1.Department of Chemistry, College of Liberal Arts and SciencesTokyo Medical and Dental UniversityChibaJapan
  2. 2.Department of Applied Biological Chemistry, Graduate School of Agricultural and Life SciencesUniversity of TokyoTokyoJapan

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