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  FTIR difference spectroscopy of the Na,K-ATPase

Stolz, M., Lewitzki, E., Thoenges, D., Mäntele, W., Barth, A., & Grell, E. (2005). FTIR difference spectroscopy of the Na,K-ATPase. Poster presented at 59th Annual Meeting of the Society of General Physiologists, Marine Biological Laboratory, Woods Hole Massachusetts.

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 Creators:
Stolz, Michael1, Author              
Lewitzki, Erwin1, Author              
Thoenges, Detlef2, Author
Mäntele, Werner2, Author
Barth, Andreas3, Author
Grell, Ernst1, Author              
Affiliations:
1Department of Biophysical Chemistry, Max Planck Institute of Biophysics, Max Planck Society, ou_2068289              
2Institute of Biophysics, University of Frankfurt am Main, Frankfurt am Main, Germany, ou_persistent22              
3Department of Biochemistry and Biophysics, The Arrhenius Laboratories for Natural Sciences, Stockholm University, Stockholm, Sweden, ou_persistent22              

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 Abstract: Changes of protein secondary structure and protein microenvironment associated with partial reactions of Na,K-ATPase have been investigated in detail by reaction-induced FTIR difference spectroscopy. Infrared absorbance changes were recorded in the region from 2000 to 950 cm−1 in H2O and D2O. Depending on medium composition, three different partial reactions have been induced by the photochemical release of ATP from different precursors (mainly NPE-caged ATP): (1) ATP binding and release, respectively, at the binding site, (2) formation of the ADP-sensitive phosphoenzyme E1P, and (3) formation of the K+-sensitive phosphoenzyme E2P. All partial reactions lead to distinct changes of the infrared spectrum that are characteristic of the adopted states. The observed band amplitudes in the amide I region of the infrared spectrum indicate that the net change of secondary structure change involves at most ∼0.3% of all amino acid residues of Na,K-ATPase. In comparison, the smallest spectral changes are found upon ATP release from the binding site, and the largest for phosphoenzyme conversion (E1P→E2P), indicating larger secondary structure changes. Phosphorylation is accompanied by the appearance of carbonyl bands at 1738 and 1709 cm−1, one of which is tentatively assigned to the phosphorylated Asp369 side chain. Spectral changes in the ranges of 1750–1700 and 1610–1400 cm−1 found upon phosphoenzyme conversion accompanied with Na+ release can be attributed to changes in hydrogen bonding, protonation state, and alkali ion coordination of carboxyl groups. The FTIR-difference spectra of Na,K-ATPase and Ca-ATPase (Barth et al. 1996. J. Biol. Chem. 271:30637–30646) differ significantly, probably due to different structural features of these two enzymes.

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Language(s): eng - English
 Dates: 2005-07-01
 Publication Status: Published in print
 Pages: 1
 Publishing info: -
 Table of Contents: -
 Rev. Type: Peer
 Identifiers: DOI: 10.1085/jgp.126.1.1a
 Degree: -

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Title: 59th Annual Meeting of the Society of General Physiologists
Place of Event: Marine Biological Laboratory, Woods Hole Massachusetts
Start-/End Date: 2005-09-06 - 2005-09-11

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Title: Journal of General Physiology
  Other : J. Gen. Physiol.
  Abbreviation : JGP
Source Genre: Journal
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Publ. Info: Rockefeller University Press
Pages: - Volume / Issue: 126 (1) Sequence Number: 70 Start / End Page: 31a - 31a Identifier: ISSN: 0022-1295
CoNE: https://pure.mpg.de/cone/journals/resource/954925413895