A C-terminal, calmodulin-like regulatory domain from the plasma membrane Ca2+-pumping ATPase.
A cDNA that encodes what appears to be the inhibitory domain of the plasma membrane calcium-pumping ATPase (Ca2+-ATPase) has been isolated by screening a lambda gt11 bovine brain cDNA library with antibodies prepared against the human erythrocyte membrane Ca2+-ATPase. This screening resulted in isolation of a bacteriophage containing a 1.5-kilobase cDNA insert encoding a 71-residue polypeptide, the remainder being a large 3' terminal noncoding region. A portion of this deduced peptide sequence was identical to that of a peptide isolated from a V8 protease digest of the human erythrocyte Ca2+-ATPase except for 1 residue. Antibodies purified by immunoabsorption to the fusion protein containing this cDNA-encoded polypeptide reacted only with those fragments of a limited trypsin digest of the human erythrocyte Ca2+-ATPase that contain the inhibitory domain. Moreover, these antibodies were able to partially stimulate basal enzyme activity and block further activation by calmodulin. The encoded polypeptide bears homology to the glutamic acid-rich regions N-terminal to the Ca2+-binding loops of calmodulin and to a lesser extent with the loops themselves. This encoded polypeptide also represents the C terminus of the Ca2+-ATPase. Portions of the isolated cDNA were homologous to the 3' noncoding region of the sarcoplasmic reticulum Ca2+-ATPase cDNA, indicating a possible mechanism for the evolution of these distinct membrane Ca2+ pumps.
Brandt
P
P
Department of Biochemistry, University of Kentucky Medical Center, Lexington 40536-0084.
Zurini
M
M
Neve
R L
RL
Rhoads
R E
RE
Vanaman
T C
TC
D000595:Amino Acid Sequence; D001483:Base Sequence; D000252:Calcium-Transporting ATPases / Q000032:analysis* / Q000235:genetics; D002147:Calmodulin / Q000378:metabolism*; D004247:DNA / Q000032:analysis; D004910:Erythrocyte Membrane / Q000201:enzymology*; D006801:Humans; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012519:Sarcoplasmic Reticulum / Q000201:enzymology; D012689:Sequence Homology, Nucleic Acid; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics
D002147:Calmodulin; D011993:Recombinant Fusion Proteins; D004247:DNA; D001616:beta-Galactosidase; D000252:Calcium-Transporting ATPases