Plasma membrane Ca(2+) pump isoform 3f is weakly stimulated by calmodulin.
Isoform 3f of the plasma membrane Ca(2+) pump is a major isoform of this pump in rat skeletal muscle. It has an unusual structure, with a short carboxyl-terminal regulatory region of only 33 residues when compared with the 77 to 124 residues found in the other isoforms. Also, whereas the regulatory regions of the other isoforms, downstream of the alternative splice, consist of two homologous groups, the sequence of 3f is not related to either group. A synthetic peptide representing the calmodulin binding domain of isoform 3f had a much lower calmodulin affinity (with a K(d) of 15 nM) than the corresponding peptide of isoform 2b (K(d) value was 0.2 nM). The characteristics of this domain were further studied by making chimeras of the 3f regulatory region with the catalytic core of isoform 4 and by making the full-length isoform 3f. Both constructs bound to calmodulin-Sepharose. The chimera was fully active without calmodulin, showing no stimulation of activity when calmodulin was added. The full-length isoform 3f was slightly activated by calmodulin. These data show that the regulatory region of isoform 3f is only a weak autoinhibitor of the enzyme, in contrast to the properties of all the other isoforms studied so far. Rather, this isoform is a special-purpose, constitutively active form of the enzyme, expressed primarily in skeletal muscle and as a minor isoform in brain.
Filoteo
A G
AG
Department of Biochemistry and Molecular Biology, Mayo Foundation, Rochester, Minnesota 55905, USA.
Enyedi
A
A
Verma
A K
AK
Elwess
N L
NL
Penniston
J T
JT
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D019556:COS Cells; D000252:Calcium-Transporting ATPases / Q000378:metabolism*; D002147:Calmodulin / Q000494:pharmacology*; D027682:Cation Transport Proteins; D002462:Cell Membrane / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D008969:Molecular Sequence Data; D018482:Muscle, Skeletal / Q000201:enzymology; D010446:Peptide Fragments / Q000378:metabolism / Q000494:pharmacology; D053499:Plasma Membrane Calcium-Transporting ATPases; D011485:Protein Binding; D051381:Rats; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D014162:Transfection
D002147:Calmodulin; D027682:Cation Transport Proteins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D053499:Plasma Membrane Calcium-Transporting ATPases; D000252:Calcium-Transporting ATPases