Photoaffinity labeling study of the interaction of calmodulin with the plasma membrane Ca2+ pump.

Details

Serval ID
serval:BIB_853ECF0ACEBD
Type
Article: article from journal or magazin.
Collection
Publications
Title
Photoaffinity labeling study of the interaction of calmodulin with the plasma membrane Ca2+ pump.
Journal
Biochemistry
Author(s)
Vorherr T., Quadroni M., Krebs J., Carafoli E.
ISSN
0006-2960
Publication state
Published
Issued date
09/1992
Peer-reviewed
Oui
Volume
31
Number
35
Pages
8245-8251
Language
english
Abstract
Bovine brain calmodulin was labeled with synthetic peptides corresponding to the calmodulin-binding domain of the erythrocyte plasma membrane Ca(2+)-ATPase. One 20-amino acid peptide and two 28-amino acid peptides were used, carrying L-4'-(1-azi-2,2,2-trifluoroethyl)phenylalanine residues in position 9 (peptides C20W* and C28W*) and position 25 (peptide C28WC*), respectively. The localization of the contact regions between calmodulin and the N- and C-terminal portions of the peptides was the aim of this study. The three peptides were N-terminally blocked with a 3H-labeled acetyl group to facilitate the identification of labeled fragments after isolation and digestion. The binding site for phenylalanine 25 was identified in the N-terminal domain of calmodulin while the phenylalanine derivative in position 9 labeled the C-terminal domain. Fluorescence studies using the dansylated N- and C-terminal halves of calmodulin and peptide C20W corresponding to the first 20 amino acids of the calmodulin-binding domain showed that only the C-terminal lobe of calmodulin had high affinity for the peptide (KD in the nanomolar range).
Keywords
Affinity Labels, Amino Acid Sequence, Animals, Binding Sites, Brain, Calcium-Transporting ATPases, Calmodulin, Cattle, Cell Membrane, Chromatography, High Pressure Liquid, Cross-Linking Reagents, Cyanogen Bromide, Endopeptidases, Erythrocyte Membrane, Kinetics, Molecular Sequence Data, Peptide Fragments
Pubmed
Web of science
Create date
24/01/2008 15:46
Last modification date
20/08/2019 14:44
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