title: The adaptors Grb10 and Grb14 are calmodulin-binding proteins.
abstract: We identified the Grb7 family members, Grb10 and Grb14, as Ca2+-dependent CaM-binding proteins using Ca2+-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.
authors.affiliation: Department of Cancer Biology, Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, Spain.
authors.forename: Irene
authors.initials: I
authors.lastname: García-Palmero
authors.affiliation: Department of Cancer Biology, Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, Spain.
authors.forename: Noemí
authors.initials: N
authors.lastname: Pompas-Veganzones
authors.affiliation: Departamento de Microbiología, Facultad de Biología, Universidad de Sevilla, Spain.
authors.forename: Eduardo
authors.initials: E
authors.lastname: Villalobo
authors.affiliation: Plate-forme de Chimie Biologique Intégrative de Strasbourg (PCBIS), UMS 3286 CNRS-Université de Strasbourg, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Gioria
authors.affiliation: Laboratoire d'Excellence Medalis, Université de Strasbourg, CNRS, LIT UMR 7200, France.
authors.forename: Jacques
authors.initials: J
authors.lastname: Haiech
authors.affiliation: Department of Cancer Biology, Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, Spain.
authors.forename: Antonio
authors.initials: A
authors.lastname: Villalobo
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D001665:Binding Sites; D020013:Calcium Signaling; D002147:Calmodulin / Q000737:chemistry / Q000378:metabolism*; D002846:Chromatography, Affinity; D017124:Conserved Sequence; D051383:GRB10 Adaptor Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051382:GRB7 Adaptor Protein / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D057809:HEK293 Cells; D006801:Humans; D007700:Kinetics; D016297:Mutagenesis, Site-Directed; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein
chemical_list: D048868:Adaptor Proteins, Signal Transducing; C000607143:CALM1 protein, human; D002147:Calmodulin; C096358:GRB10 protein, human; C099655:GRB14 protein, human; C495955:GRB7 protein, human; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; D051382:GRB7 Adaptor Protein; D051383:GRB10 Adaptor Protein
keywords: Grb10; Grb14; Grb7; adaptor proteins; calmodulin; calmodulin-binding sites