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The adaptors Grb10 and Grb14 are calmodulin‐binding proteins
Author(s) -
GarcíaPalmero Irene,
PompasVeganzones Noemí,
Villalobo Eduardo,
Gioria Sophie,
Haiech Jacques,
Villalobo Antonio
Publication year - 2017
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1002/1873-3468.12623
Subject(s) - calmodulin , chemistry , binding site , plasma protein binding , biochemistry , dna binding protein , binding constant , transcription factor , enzyme , gene
We identified the Grb7 family members, Grb10 and Grb14, as Ca 2+ ‐dependent CaM‐binding proteins using Ca 2+ ‐dependent CaM‐affinity chromatography as we previously did with Grb7. The potential CaM‐binding sites were identified and experimentally tested using fluorescent‐labeled peptides corresponding to these sites. The apparent affinity constant of these peptides for CaM, and the minimum number of calcium ions bound to CaM that are required for effective binding to these peptides were also determined. We prepared deletion mutants of the three adaptor proteins lacking the identified sites and determined that they lost or strongly diminished their CaM‐binding capacity following the sequence Grb7 > > Grb14 > Grb10. More than one CaM‐binding site and/or accessory CaM‐binding sites appear to exist in Grb10 and Grb14, as compared to a single one present in Grb7.