help button home button Biophys. J.
HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS

Biophysical Journal 64: 1232-1242 (1993)
© 1993 the Biophysical Society

This Article
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Yu, X
Right arrow Articles by Inesi, G
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Yu, X
Right arrow Articles by Inesi, G

H+ countertransport and electrogenicity of the sarcoplasmic reticulum Ca2+ pump in reconstituted proteoliposomes.

X Yu, S Carroll, J L Rigaud and G Inesi

Department of Biological Chemistry, School of Medicine, University of Maryland, Baltimore 21201.

ABSTRACT

The Ca2+ transport adenosine triphosphatase of sarcoplasmic reticulum was reconstituted in unilamellar liposomes prepared by reverse-phase evaporation. The size of the resulting proteoliposomes was similar to that of native sarcoplasmic reticulum vesicles, but their protein content was much lower, with a protein/lipid ratio (wt/wt) of 1:40-160, as compared with 1:1 in the native membrane. The proteoliposomes sustained adenosine triphosphate-dependent Ca2+ uptake at rates proportional to the protein content (1-2 mumol Ca2+/mg protein/min), reaching asymptotic levels corresponding to a lumenal calcium concentration of 10-20 mM. The low permeability of the proteoliposomes permitted direct demonstration of Ca2+/H+ countertransport and electrogenicity by parallel measurements in the same experimental system. Countertransport of one H+ per one Ca2+ was demonstrated, and inhibition of the Ca2+ pump by lumenal alkalinization was relieved by the H+ ionophore carbonyl cyanide p-(trifluoromethoxy)phenylhydrazone. Consistent with the countertransport stoichiometry, net positive charge displacement was produced by Ca2+ transport, as revealed by a rapid oxonol VI absorption rise. The initial rise and the following steady-state level of oxonol absorption were highest when SO4(2-) was the prevalent anion and lowest in the presence of the lipophilic anion SCN-. The influence of anions was attributed to potential driven counterion compensation. The absorption rise was rapidly collapsed by addition of valinomycin in the presence of K+. Experimentation with Ca2+ and H+ ionophores was consistent with a primary role of Ca2+ and H+ in net charge displacement. The estimated value of the steady-state electrical potential observed under optimal conditions was approximately 50 mV and was accounted for by the estimated charge transfer associated with Ca2+ and H+ countertransport under the same conditions.




This article has been cited by other articles:


Home page
 Am. J. Physiol. Cell Physiol.Home page
R. J. Scheibe, K. Mundhenk, T. Becker, J. Hallerdei, A. Waheed, G. N. Shah, W. S. Sly, G. Gros, and P. Wetzel
Carbonic anhydrases IV and IX: subcellular localization and functional role in mouse skeletal muscle
Am J Physiol Cell Physiol, February 1, 2008; 294(2): C402 - C412.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
A. Fibich, K. Janko, and H.-J. Apell
Kinetics of Proton Binding to the Sarcoplasmic Reticulum Ca-ATPase in the E1 State
Biophys. J., November 1, 2007; 93(9): 3092 - 3104.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
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]

Home page
 Biophys. JHome page
C. Peinelt and H.-J. Apell
Kinetics of Ca2+ Binding to the SR Ca-ATPase in the E1 State
Biophys. J., October 1, 2005; 89(4): 2427 - 2433.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
A. S. Fraysse, A. L. B. Moller, L. R. Poulsen, B. Wollenweber, M. J. Buch-Pedersen, and M. G. Palmgren
A Systematic Mutagenesis Study of Ile-282 in Transmembrane Segment M4 of the Plasma Membrane H+-ATPase
J. Biol. Chem., June 10, 2005; 280(23): 21785 - 21790.
[Abstract] [Full Text] [PDF]

Home page
 ScienceHome page
C. Olesen, T. L.-M. Sorensen, R. C. Nielsen, J. V. Moller, and P. Nissen
Dephosphorylation of the Calcium Pump Coupled to Counterion Occlusion
Science, December 24, 2004; 306(5705): 2251 - 2255.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
F. T. Buoninsegni, G. Bartolommei, M. R. Moncelli, G. Inesi, and R. Guidelli
Time-Resolved Charge Translocation by Sarcoplasmic Reticulum Ca-ATPase Measured on a Solid Supported Membrane
Biophys. J., June 1, 2004; 86(6): 3671 - 3686.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
C. Peinelt and H.-J. Apell
Time-Resolved Charge Movements in the Sarcoplasmatic Reticulum Ca-ATPase
Biophys. J., February 1, 2004; 86(2): 815 - 824.
[Abstract] [Full Text] [PDF]

Home page
 Biophys. JHome page
N. Reuter, K. Hinsen, and J.-J. Lacapere
Transconformations of the SERCA1 Ca-ATPase: A Normal Mode Study
Biophys. J., October 1, 2003; 85(4): 2186 - 2197.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. V. Moller, G. Lenoir, C. Marchand, C. Montigny, M. le Maire, C. Toyoshima, B. S. Juul, and P. Champeil
Calcium Transport by Sarcoplasmic Reticulum Ca2+-ATPase. ROLE OF THE A DOMAIN AND ITS C-TERMINAL LINK WITH THE TRANSMEMBRANE REGION
J. Biol. Chem., October 4, 2002; 277(41): 38647 - 38659.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Cell Physiol.Home page
P. Wetzel, T. Kleinke, S. Papadopoulos, and G. Gros
Inhibition of muscle carbonic anhydrase slows the Ca2+ transient in rat skeletal muscle fibers
Am J Physiol Cell Physiol, October 1, 2002; 283(4): C1242 - C1253.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. M. Salvador, G. Inesi, J.-L. Rigaud, and A. M. Mata
Ca2+ Transport by Reconstituted Synaptosomal ATPase Is Associated with H+ Countertransport and Net Charge Displacement
J. Biol. Chem., July 17, 1998; 273(29): 18230 - 18234.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
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]

Home page
 J. Biol. Chem.Home page
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]

Home page
 J. Biol. Chem.Home page
X. Yu and G. Inesi
Variable Stoichiometric Efficiency of Ca[IMAGE] and Sr[IMAGE] Transport by the Sarcoplasmic Reticulum ATPase
J. Biol. Chem., March 3, 1995; 270(9): 4361 - 4367.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
J. P. Andersen and J. P. Andersen
Functional Consequences of Alterations to Amino Acids at the M5S5 Boundary of the Ca[IMAGE]-ATPase of Sarcoplasmic Reticulum
J. Biol. Chem., January 13, 1995; 270(2): 908 - 914.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
T. L.-M. Sorensen and J. P. Andersen
Importance of Stalk Segment S5 for Intramolecular Communication in the Sarcoplasmic Reticulum Ca2+-ATPase
J. Biol. Chem., September 8, 2000; 275(37): 28954 - 28961.
[Abstract] [Full Text] [PDF]


HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS
Copyright © 1993 by the Biophysical Society.