Organization of functional domains in the docking protein p130Cas.
The docking protein p130Cas becomes phosphorylated upon cell adhesion to extracellular matrix proteins, and is thought to play an essential role in cell transformation. Cas transmits signals through interactions with the Src-homology 3 (SH3) and Src-homology 2 domains of FAK or v-Crk signaling molecules, or with 14-3-3 protein, as well as phosphatases PTP1B and PTP-PEST. The large (130kDa), multi-domain Cas molecule contains an SH3 domain, a Src-binding domain, a serine-rich protein interaction region, and a C-terminal region that participates in protein interactions implicated in antiestrogen resistance in breast cancer. In this study, as part of a long-term goal to examine the protein interactions of Cas by X-ray crystallography and nuclear magnetic resonance spectroscopy, molecular constructs were designed to express two adjacent domains, the serine-rich domain and the Src-binding domain, that each participate in intermolecular contacts dependent on protein phosphorylation. The protein products are soluble, homogeneous, monodisperse, and highly suitable for structural studies to define the role of Cas in integrin-mediated cell signaling.
Nasertorabi
Fariborz
F
Cancer Center, The Burnham Institute, La Jolla, CA 92037, USA.
Garcia-Guzman
Miguel
M
Briknarová
Klára
K
Larsen
Elise
E
Havert
Marnie L
ML
Vuori
Kristiina
K
Ely
Kathryn R
KR
D048948:14-3-3 Proteins / Q000737:chemistry; D000818:Animals; D002471:Cell Transformation, Neoplastic; D002942:Circular Dichroism; D050719:Crk-Associated Substrate Protein; D018360:Crystallography, X-Ray; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000378:metabolism; D016023:Integrins; D008027:Light; D009682:Magnetic Resonance Spectroscopy; D010766:Phosphorylation; D011485:Protein Binding; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12; D017027:Protein Tyrosine Phosphatases / Q000737:chemistry / Q000378:metabolism; D011506:Proteins / Q000737:chemistry* / Q000378:metabolism; D051381:Rats; D050717:Retinoblastoma-Like Protein p130; D012542:Scattering, Radiation; D012694:Serine / Q000737:chemistry; D015398:Signal Transduction; D018909:src Homology Domains; D019061:src-Family Kinases / Q000378:metabolism
D048948:14-3-3 Proteins; C493843:Bcar1 protein, rat; D050719:Crk-Associated Substrate Protein; D016023:Integrins; D011506:Proteins; D050717:Retinoblastoma-Like Protein p130; D012694:Serine; D005982:Glutathione Transferase; D019061:src-Family Kinases; D054562:Protein Tyrosine Phosphatase, Non-Receptor Type 1; D054594:Protein Tyrosine Phosphatase, Non-Receptor Type 12; D017027:Protein Tyrosine Phosphatases; C516610:Ptpn1 protein, rat; C516740:Ptpn12 protein, rat