Biophysical Journal 71: 2970-2983 (1996)
© 1996 the Biophysical Society

This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Troullier, A Articles by Dupont, Y Search for Related Content PubMed PubMed Citation Articles by Troullier, A Articles by Dupont, Y

A time-resolved Fourier transformed infrared difference spectroscopy study of the sarcoplasmic reticulum Ca(2+)-ATPase: kinetics of the high-affinity calcium binding at low temperature.

C.E.A., Laboratoire de Biophysique Moléculaire et Cellulaire, URA CNRS 520, Département de Biologie Moléculaire et Structurale, Grenoble, France.

ABSTRACT

We have used time-resolved Fourier transformed infrared difference spectroscopy to characterize the amplitude, frequency, and kinetics of the absorbance changes induced in the infrared (IR) spectrum of sarcoplasmic reticulum Ca(2+)-ATPase by calcium binding at the high-affinity transport sites. 1-(2-Nitro-4,5-dimethoxyphenyl)-N,N,N',N'-tetrakis [(oxycarbonyl)methyl]-1,2-ethanediamine (DM-nitrophen) was used as a caged-calcium compound to trigger the release of calcium in the IR samples. Calcium binding to Ca(2+)-ATPase induces the appearance of spectral bands in difference spectra that are all absent in the presence of the inhibitor thapsigargin. Spectral bands above 1700 cm-1 indicate that glutamic and/or aspartic acid side chains are deprotonated upon calcium binding, whereas other bands may be induced by reactions of asparagine, glutamine, and tyrosine residues. Some of the bands appearing in the 1690-1610 cm-1 region arise from modifications of peptide backbone carbonyl groups. The band at 1653 cm-1 is a candidate for a change in an alpha-helix, whereas other bands could arise from modifications of random, turn, or beta-sheet structures or from main-chain carbonyl groups playing the role of calcium ligands. Only a few residues are involved in secondary structure changes. The kinetic evolution of these bands was recorded at low temperature (-9 degrees C). All bands exhibited a monophasic kinetics of rate constant 0.026 s-1, which is compatible with that measured in previous study at the same temperature in a suspension of sarcoplasmic reticulum vesicles by intrinsic fluorescence of Ca(2+)-ATPase.

This article has been cited by other articles:

				J. Andersson, K. Hauser, E.-L. Karjalainen, and A. Barth Protonation and Hydrogen Bonding of Ca2+ Site Residues in the E2P Phosphoenzyme Intermediate of Sarcoplasmic Reticulum Ca2+-ATPase Studied by a Combination of Infrared Spectroscopy and Electrostatic Calculations Biophys. J., January 15, 2008; 94(2): 600 - 611. [Abstract] [Full Text] [PDF]

				K. Hauser and A. Barth Side-Chain Protonation and Mobility in the Sarcoplasmic Reticulum Ca2+-ATPase: Implications for Proton Countertransport and Ca2+ Release Biophys. J., November 1, 2007; 93(9): 3259 - 3270. [Abstract] [Full Text] [PDF]

				B. A. Barry, C. Hicks, A. De Riso, and D. L. Jenson Calcium Ligation in Photosystem II under Inhibiting Conditions Biophys. J., July 1, 2005; 89(1): 393 - 401. [Abstract] [Full Text] [PDF]

				M. Liu, E.-L. Karjalainen, and A. Barth Use of Helper Enzymes for ADP Removal in Infrared Spectroscopic Experiments: Application to Ca2+-ATPase Biophys. J., May 1, 2005; 88(5): 3615 - 3624. [Abstract] [Full Text] [PDF]

				L. Zhang, R. Buchet, and G. Azzar Phosphate Binding in the Active Site of Alkaline Phosphatase and the Interactions of 2-Nitrosoacetophenone with Alkaline Phosphatase-Induced Small Structural Changes Biophys. J., June 1, 2004; 86(6): 3873 - 3881. [Abstract] [Full Text] [PDF]

				A. Barth Phosphoenzyme Conversion of the Sarcoplasmic Reticulum Ca2+-ATPase. MOLECULAR INTERPRETATION OF INFRARED DIFFERENCE SPECTRA J. Biol. Chem., August 6, 1999; 274(32): 22170 - 22175. [Abstract] [Full Text] [PDF]

				D. H. MacLennan, W. J. Rice, and N. M. Green The Mechanism of Ca2+ Transport by Sarco(Endo)plasmic Reticulum Ca2+-ATPases J. Biol. Chem., November 14, 1997; 272(46): 28815 - 28818. [Full Text] [PDF]

				A. Barth, W. Kreutz, and W. Mantele Ca2+ Release from the Phosphorylated and the Unphosphorylated Sarcoplasmic Reticulum Ca2+ ATPase Results in Parallel Structural Changes. AN INFRARED SPECTROSCOPIC STUDY J. Biol. Chem., October 10, 1997; 272(41): 25507 - 25510. [Abstract] [Full Text] [PDF]