The effect of dicyclohexycarbodiimide and cyclopiazonic acid on the difference FTIR spectra of sarcoplasmic reticulum induced by photolysis of caged-ATP and caged-Ca2+

Rene Buchet, I. Jóna, Anthony Martonosi

Research output: Contribution to journalArticle

23 Citations (Scopus)

Abstract

The photochemical release of Ca2+ from caged-Ca2+ in the absence of ATP, and the release of ATP from caged-ATP in the presence of Ca2+ induce characteristic difference FTIR spectra on rabbit sarcoplasmic reticulum that are related to the formation of Ca2E1 and E ∼ P intermediates of the Ca2+-ATPase, respectively. Dicyclohexylcarbodiimide (10 nmol/mg protein) abolished both the Ca2+-and ATP-induced difference FTIR spectra parallel with inhibition of ATPase activity. Cyclopiazonic acid (50 nmol/mg protein) inhibited the Ca2+-induced difference spectrum measured in the absence of ATP, but had no significant effect on the ATP-induced difference spectrum measured in the presence of 1 mM Ca2+. The dog kidney Na+,K+-ATPase did not give significant difference spectrum after photolysis of caged-ATP in Ca2+-free media containing 90 mM Na+ and 10 mM K+, with or without ouabain. We propose that both the Ca2+ and the ATP-induced difference FTIR spectra of the Ca2+-ATPase reflect the occupancy of the high-affinity Ca2+ transport site of the enzyme.

Original languageEnglish
Pages (from-to)207-214
Number of pages8
JournalBBA - Biomembranes
Volume1104
Issue number1
DOIs
Publication statusPublished - Feb 17 1992

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Photolysis
Sarcoplasmic Reticulum
Fourier Transform Infrared Spectroscopy
Adenosine Triphosphate
Calcium-Transporting ATPases
Adenosine Triphosphatases
Dicyclohexylcarbodiimide
Adenosinetriphosphate
Ouabain
Proteins
cyclopiazonic acid
P(3)-1-(2-nitro)phenylethyladenosine 5'-triphosphate
Dogs
Rabbits
Kidney
Enzymes

Keywords

  • ATPase, Ca-
  • Caged ATP
  • Caged calcium
  • FTIR
  • Photolysis
  • Sarcoplasmic reticulum

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Cell Biology

Cite this

The effect of dicyclohexycarbodiimide and cyclopiazonic acid on the difference FTIR spectra of sarcoplasmic reticulum induced by photolysis of caged-ATP and caged-Ca2+. / Buchet, Rene; Jóna, I.; Martonosi, Anthony.

In: BBA - Biomembranes, Vol. 1104, No. 1, 17.02.1992, p. 207-214.

Research output: Contribution to journalArticle

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