Expression of the sarcoplasmic reticulum Ca2+‐ATPase in yeast

FEBS Letters

Falson, Pierre³  Centeno, Francisco³  Moutin, Marie-Jo²  Villalba, Jose Manuel⁶  le Maire, Marc³  Palmgren, Michael Gjedde⁴  Deschamps, Stéphane³  Dupont, Yves²  Anger, Marielle⁵  Møller, Jesper Vuust¹  Lompré, Anne-Marie⁵ 

[1] Danish Biomembrane Research Centre, Institute of Biophysics, University of Aarhus, DK-8000 Aarhus C, Denmark;Laboratoire de Biophysique Moléculaire et Cellulaire, URA 520 CNRS, CEA, DBMS/CEG, 17 rue des Martyrs, 85X, F-38041 Grenoble, France;Section de Biophysique des Protéines et des Membranes, Département de Biologie Cellulaire et Moléculaire, CEA et CNRS URA 1290, CE Saclay, 91191 Gif sur Yvette, France;Department of Plant Biology, The Royal Veterinary and Agricultural University, Thorvaldsensvej 40, DK-1871 Frederiksberg C, Copenhagen, Denmark;URA CNRS 1131, Université de Paris-Sud, Centre d'Orsay, 91405 Orsay, France;Departamento de Biologia Celular, Facultad de Ciencias, 14004 Córdoba, Spain

关键词: SR;    sarcoplasmic reticulum;    SERCA;    sarco(endo)-plasmic reticulum Ca2+-ATPase;    PM;    plasma membrane;    C12E8;    octa-ethyleneglycol monododecylether;    TES;    2-{[2-hydroxy-1;    1-bis-(hydroxymethyl)ethyl]amino} ethanesulfonic acid;    pBS;    plasmid bluescript (Stratagene);    MOPS;    3-[N-morpholino]propanosulfonic acid;    EGTA;    ethyleneglycol-bis-(β-amino-ethyl ether) N;    N′-tetra-acetic acid;    pAb;    polyclonal antibody;    SDS;    sodium dodecyl sulfate;    PAGE;    polyacrylamide gel electrophoresis;    PVDF;    polyvinylidene difluoride;    TCA trichloroacetic acid;    FITC;    fluorescein isothiocyanate;    ATP;    adenosine-5′-triposphate;    NADH;    nicotinamide adenine dinucleotide;    reduced form;    RT-PCR;    reverse transcriptase-polymease chain reaction;    DAPI;    4′;    6-diamidino-2-phenylindole;   

DOI  :  10.1016/0014-5793(94)01104-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We describe here an easy system for the production of mg amounts of the rabbit Ca²⁺-ATPase SERCA 1a in the yeast S. cerevisiae. The protein is present in several membranes, including the plasma membrane of the yeast, in a native conformation. It can be purified by immunoprecipitation and can be phosphorylated from ATP in a Ca²⁺-dependent manner. Using a temperature-sensitive secretion mutant strain, the fully active protein can also be obtained in secretory vesicles.

【 授权许可】

Unknown