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Title: Interaction of calmodulin with the calmodulin binding domain of the plasma membrane Ca sup 2+ pump

Abstract

Peptides corresponding to the calmodulin binding domain of the plasma membrane Ca{sup 2+} pump 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 Ca{sup 2+} binding site in the pump was also synthesized. Circular dichroism experiments have shownmore » that it interacted with calmodulin binding domain, supporting the suggestion that the latter, or a portion of it, may act as a natural inhibitor of the pump.« less

Authors:
; ; ;  [1]; ;  [2];  [3]
  1. (Swiss Federal Institute of Technology, Zurich (Switzerland))
  2. (Mayo Clinic, Rochester, MN (USA))
  3. (National Institute of Haematology and Blood Transfusion, Budapest (Hungary))
Publication Date:
OSTI Identifier:
6718575
Alternate Identifier(s):
OSTI ID: 6718575
Resource Type:
Journal Article
Journal Name:
Biochemistry; (USA)
Additional Journal Information:
Journal Volume: 29:2; Journal ID: ISSN 0006-2960
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; CALMODULIN; BIOLOGICAL FUNCTIONS; PORINS; BIOCHEMICAL REACTION KINETICS; AMINO ACID SEQUENCE; CHEMICAL SHIFT; MAGNETIC CIRCULAR DICHROISM; NUCLEAR MAGNETIC RESONANCE; OVERHAUSER EFFECT; TRYPTOPHAN; AMINO ACIDS; AROMATICS; AZAARENES; AZOLES; CARBOXYLIC ACIDS; DICHROISM; FUNCTIONS; HETEROCYCLIC ACIDS; HETEROCYCLIC COMPOUNDS; INDOLES; KINETICS; MAGNETIC RESONANCE; MEMBRANE PROTEINS; MOLECULAR STRUCTURE; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC NITROGEN COMPOUNDS; PROTEINS; PYRROLES; REACTION KINETICS; RESONANCE 550201* -- Biochemistry-- Tracer Techniques

Citation Formats

Vorherr, T., James, P., Krebs, J., Carafoli, E., McCormick, D.J., Penniston, J.T., and Enyedi, A. Interaction of calmodulin with the calmodulin binding domain of the plasma membrane Ca sup 2+ pump. United States: N. p., 1990. Web. doi:10.1021/bi00454a008.
Vorherr, T., James, P., Krebs, J., Carafoli, E., McCormick, D.J., Penniston, J.T., & Enyedi, A. Interaction of calmodulin with the calmodulin binding domain of the plasma membrane Ca sup 2+ pump. United States. doi:10.1021/bi00454a008.
Vorherr, T., James, P., Krebs, J., Carafoli, E., McCormick, D.J., Penniston, J.T., and Enyedi, A. Tue . "Interaction of calmodulin with the calmodulin binding domain of the plasma membrane Ca sup 2+ pump". United States. doi:10.1021/bi00454a008.
@article{osti_6718575,
title = {Interaction of calmodulin with the calmodulin binding domain of the plasma membrane Ca sup 2+ pump},
author = {Vorherr, T. and James, P. and Krebs, J. and Carafoli, E. and McCormick, D.J. and Penniston, J.T. and Enyedi, A.},
abstractNote = {Peptides corresponding to the calmodulin binding domain of the plasma membrane Ca{sup 2+} pump 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 Ca{sup 2+} binding site in the pump was also synthesized. Circular dichroism experiments have shown that it interacted with calmodulin binding domain, supporting the suggestion that the latter, or a portion of it, may act as a natural inhibitor of the pump.},
doi = {10.1021/bi00454a008},
journal = {Biochemistry; (USA)},
issn = {0006-2960},
number = ,
volume = 29:2,
place = {United States},
year = {1990},
month = {1}
}