The serine-rich domain from Crk-associated substrate (p130cas) is a four-helix bundle.
p130(cas) (Crk-associated substrate) is a docking protein that is involved in assembly of focal adhesions and concomitant cellular signaling. It plays a role in physiological regulation of cell adhesion, migration, survival, and proliferation, as well as in oncogenic transformation. The molecule consists of multiple protein-protein interaction motifs, including a serine-rich region that is positioned between Crk and Src-binding sites. This study reports the first structure of a functional domain of Cas. The solution structure of the serine-rich region has been determined by NMR spectroscopy, demonstrating that this is a stable domain that folds as a four-helix bundle, a protein-interaction motif. The serine-rich region bears strong structural similarity to four-helix bundles found in other adhesion components like focal adhesion kinase, alpha-catenin, or vinculin. Potential sites for phosphorylation and interaction with the 14-3-3 family of cellular regulators are identified in the domain and characterized by site-directed mutagenesis and binding assays. Mapping the degree of amino acid conservation onto the molecular surface reveals a patch of invariant residues near the C terminus of the bundle, which may represent a previously unidentified site for protein interaction.
Briknarová
Klára
K
Burnham Institute, 10901 N. Torrey Pines Road, La Jolla, CA 92037, USA.
Nasertorabi
Fariborz
F
Havert
Marnie L
ML
Eggleston
Ericka
E
Hoyt
David W
DW
Li
Chenglong
C
Olson
Arthur J
AJ
Vuori
Kristiina
K
Ely
Kathryn R
KR
D048948:14-3-3 Proteins / Q000737:chemistry; D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002448:Cell Adhesion; D002465:Cell Movement; D049109:Cell Proliferation; D002470:Cell Survival; D002471:Cell Transformation, Neoplastic; D050719:Crk-Associated Substrate Protein; D003598:Cytoskeletal Proteins / Q000737:chemistry; D006801:Humans; D009682:Magnetic Resonance Spectroscopy; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D009154:Mutation; D010455:Peptides / Q000737:chemistry; D010766:Phosphorylation; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D011506:Proteins / Q000737:chemistry / Q000502:physiology*; D051381:Rats; D050717:Retinoblastoma-Like Protein p130; D017386:Sequence Homology, Amino Acid; D012694:Serine / Q000737:chemistry*; D015398:Signal Transduction; D016596:Vinculin / Q000737:chemistry; D051178:alpha Catenin
D048948:14-3-3 Proteins; C493841:BCAR1 protein, human; C493842:Bcar1 protein, mouse; C493843:Bcar1 protein, rat; C495273:CTNNA1 protein, human; D050719:Crk-Associated Substrate Protein; C495272:Ctnna1 protein, mouse; D003598:Cytoskeletal Proteins; D010455:Peptides; D011506:Proteins; D050717:Retinoblastoma-Like Protein p130; D051178:alpha Catenin; D016596:Vinculin; D012694:Serine