Functional Co-expression of the canine cardiac Ca2+ pump and phospholamban in Spodoptera frugiperda (Sf21) cells reveals new insights on ATPase regulation.

The Journal of Biological Chemistry
J M Autry, L R Jones

Abstract

The utility of the baculovirus cell expression system for investigating Ca2+-ATPase and phospholamban regulatory interactions was examined. cDNA encoding the canine cardiac sarco(endo)plasmic Ca2+-ATPase pump (SERCA2a) was cloned for the first time and expressed in the presence and absence of phospholamban in Spodoptera frugiperda (Sf21) insect cells. The recombinant Ca2+ pump was produced in high yield, contributing 20% of the total membrane protein in Sf21 microsomes. At least 70% of the expressed pumps were active. Co-expression of wild-type, pentameric phospholamban with the Ca2+-ATPase decreased the apparent affinity of the ATPase for Ca2+, but had no effect on the maximum velocity of the enzyme, similar to phospholamban's action in cardiac sarcoplasmic reticulum vesicles. To investigate the importance of the oligomeric structure of phospholamban in ATPase regulation, SERCA2a was co-expressed with a monomeric mutant of phospholamban, in which leucine residue 37 was changed to alanine. Surprisingly, monomeric phospholamban suppressed SERCA2a Ca2+ affinity more strongly than did wild-type phospholamban, demonstrating that the pentamer is not essential for Ca2+ pump inhibition and that the monomer is the more active species. ...Continue Reading

References

Jun 15, 1992·Biochimica Et Biophysica Acta·A M GambelD R Menick
Jun 15, 1989·The Biochemical Journal·J A EggermontR Casteels
May 1, 1989·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·M Tada, M Kadoma
May 1, 1988·Proceedings of the National Academy of Sciences of the United States of America·K Maruyama, D H MacLennan
Jul 23, 1982·Molecular and Cellular Biochemistry·M TadaF Ohmori
May 1, 1995·Biophysical Journal·J C VossD D Thomas
May 10, 1995·Molecular and Cellular Biochemistry·J M HarrerE G Kranias
Apr 21, 1995·The Journal of Biological Chemistry·L G ReddyD L Stokes
May 12, 1995·Journal of Molecular Biology·I T ArkinS O Smith
Jul 1, 1994·Biophysical Journal·J VossD D Thomas
Oct 31, 1994·FEBS Letters·F CentenoP Falson
Sep 6, 1996·The Journal of Biological Chemistry·Y KimuraD H MacLennan

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Citations

Dec 22, 1999·Archives of Biochemistry and Biophysics·J MahaneyT Muchnok
Feb 10, 2004·Protein Expression and Purification·Jason R WaggonerJames E Mahaney
Dec 5, 2003·Journal of Molecular and Cellular Cardiology·Raúl J DomenechPilar Macho
Jan 25, 2003·Cell Calcium·F WuytackL Missiaen
Aug 26, 1998·Journal of the American College of Cardiology·B S CainA H Harken
Feb 20, 2002·Trends in Cardiovascular Medicine·Dirk Hagemann, Rui-Ping Xiao
Feb 28, 2004·Trends in Cardiovascular Medicine·M Tada, T Toyofuku
Aug 22, 2000·European Journal of Biochemistry·D H MacLennan
Dec 2, 2009·The Journal of Biological Chemistry·Zhenhui ChenLarry R Jones
Jul 9, 2011·The Journal of Biological Chemistry·Joseph M AutryDavid D Thomas
Aug 8, 2013·Antioxidants & Redox Signaling·Vidhya SivakumaranNazareno Paolocci
Dec 22, 1999·Annals of the New York Academy of Sciences·D H MacLennanT Toyofuku
Dec 22, 1999·Annals of the New York Academy of Sciences·J M Autry, L R Jones
Dec 22, 1999·Annals of the New York Academy of Sciences·D D ThomasJ Stamm
Dec 22, 1999·Annals of the New York Academy of Sciences·E LoukianovM Periasamy
Jan 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Chikashi ToyoshimaDavid H MacLennan
Sep 28, 2004·Proceedings of the National Academy of Sciences of the United States of America·Christine B KarimDavid D Thomas
Apr 15, 2003·Proceedings of the National Academy of Sciences of the United States of America·Michio AsahiDavid H MacLennan
Dec 13, 2000·Journal of Molecular and Cellular Cardiology·A G Brittsan, E G Kranias
Feb 13, 2001·Journal of Molecular and Cellular Cardiology·L ZhangL R Jones
Sep 21, 2010·Biochemical and Biophysical Research Communications·Eleri HughesDavid A Middleton
Jan 7, 2006·The Journal of Investigative Dermatology·Eliane J MüllerMaja M Suter
Apr 20, 2007·Journal of Molecular and Cellular Cardiology·Poornima BhupathyMuthu Periasamy
Mar 28, 2007·Heart Failure Clinics·Jason R Waggoner, Evangelia G Kranias
Sep 1, 2004·Biochemical and Biophysical Research Communications·Kobra HaghighiEvangelia G Kranias
Apr 1, 2006·Journal of Molecular Biology·Christine B KarimDavid D Thomas
Mar 8, 2006·Journal of Molecular Biology·Jarrod J BuffyGianluigi Veglia
Sep 18, 2002·Journal of Molecular and Cellular Cardiology·David H MacLennanChulHee Kang
Sep 11, 2012·Biochimica Et Biophysica Acta·Maryam Sayadi, Michael Feig

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