Expression of the sarcoplasmic reticulum Ca(2+)-ATPase in yeast.
We describe here an easy system for the production of mg amounts of the rabbit Ca(2+)-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 Ca(2+)-dependent manner. Using a temperature-sensitive secretion mutant strain, the fully active protein can also be obtained in secretory vesicles.
Département de Biologie Cellulaire et Moléculaire, CEA, Gif sur Yvette, France.
F
F
Centeno
S
S
Deschamps
A M
AM
Lompré
M
M
Anger
M J
MJ
Moutin
Y
Y
Dupont
M G
MG
Palmgren
J M
JM
Villalba
J V
JV
Møller
P
P
Falson
D000818:Animals; D000252:Calcium-Transporting ATPases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary; D015870:Gene Expression; D015723:Gene Library; D008861:Microsomes / Q000378:metabolism; D018482:Muscle, Skeletal / Q000737:chemistry; D011817:Rabbits; D012441:Saccharomyces cerevisiae / Q000201:enzymology / Q000235:genetics*; D012519:Sarcoplasmic Reticulum / Q000201:enzymology*; D013696:Temperature
D018076:DNA, Complementary; D000252:Calcium-Transporting ATPases