TY - JOUR
T1 - Interaction of Calmodulin with the Calmodulin Binding Domain of the Plasma Membrane Ca2+ Pump
AU - Vorherr, Thomas
AU - James, Peter
AU - Krebs, Joachim
AU - Enyedi, Agnes
AU - McCormick, Daniel J.
AU - Penniston, John T.
AU - Carafoli, Ernesto
AU - Vorherr, Thomas
AU - James, Peter
AU - Krebs, Joachim
AU - McCormick, Daniel J.
AU - Penniston, John T.
AU - Carafoli, Ernesto
AU - Vorherr, Thomas
AU - James, Peter
AU - Krebs, Joachim
AU - McCormick, Daniel J.
AU - Penniston, John T.
AU - Carafoli, Ernesto
PY - 1990/1/1
Y1 - 1990/1/1
N2 - Peptides corresponding to the calmodulin binding domain of the plasma membrane Ca2+ pump (James et al., 1988) were synthesized, and their interaction with calmodulin was studied with circular dichroism, infrared spectroscopy, nuclear magnetic resonance, and fluorescence techniques. They corresponded to the complete calmodulin binding domain (28 residues), to its first 15 or 20 amino acids, and to its C-terminal 14 amino acids. The first three peptides interacted with calmodulin. The K value was similar to that of the intact enzyme in the 28 and 20 amino acid peptides, but increased substantially in the shorter 15 amino acid peptide. The 14 amino acid peptide corresponding to the C-terminal portion of the domain failed to bind calmodulin. 2D NMR experiments on the 20 amino acid peptides have indicated that the interaction occurred with the C-terminal half of calmodulin. A tryptophan that is conserved in most calmodulin binding domains of proteins was replaced by other amino acids, giving rise to modified peptides which had lower affinity for calmodulin. An 18 amino acid peptide corresponding to an acidic sequence immediately N-terminal to the calmodulin binding domain which is likely to be a Ca2+ binding site in the pump was also synthesized. Circular dichroism experiments have shown that it interacted with the calmodulin binding domain, supporting the suggestion (Benaim et al., 1984) that the latter, or a portion of it, may act as a natural inhibitor of the pump.
AB - Peptides corresponding to the calmodulin binding domain of the plasma membrane Ca2+ pump (James et al., 1988) were synthesized, and their interaction with calmodulin was studied with circular dichroism, infrared spectroscopy, nuclear magnetic resonance, and fluorescence techniques. They corresponded to the complete calmodulin binding domain (28 residues), to its first 15 or 20 amino acids, and to its C-terminal 14 amino acids. The first three peptides interacted with calmodulin. The K value was similar to that of the intact enzyme in the 28 and 20 amino acid peptides, but increased substantially in the shorter 15 amino acid peptide. The 14 amino acid peptide corresponding to the C-terminal portion of the domain failed to bind calmodulin. 2D NMR experiments on the 20 amino acid peptides have indicated that the interaction occurred with the C-terminal half of calmodulin. A tryptophan that is conserved in most calmodulin binding domains of proteins was replaced by other amino acids, giving rise to modified peptides which had lower affinity for calmodulin. An 18 amino acid peptide corresponding to an acidic sequence immediately N-terminal to the calmodulin binding domain which is likely to be a Ca2+ binding site in the pump was also synthesized. Circular dichroism experiments have shown that it interacted with the calmodulin binding domain, supporting the suggestion (Benaim et al., 1984) that the latter, or a portion of it, may act as a natural inhibitor of the pump.
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U2 - 10.1021/bi00454a008
DO - 10.1021/bi00454a008
M3 - Article
C2 - 2154244
AN - SCOPUS:0025165049
SN - 0006-2960
VL - 29
SP - 355
EP - 365
JO - Biochemistry
JF - Biochemistry
IS - 2
ER -