SH3-mediated targeting of Wrch1/RhoU by multiple adaptor proteins
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Sarah L. Risse
Abstract
Wrch1/RhoU is an atypical member of the Rho family. A major structural difference is the extended N-terminus of Wrch1 (nWrch1) containing three putative SH3 domain-binding motifs whose specificities are unknown. To define the impact of this extended region on coupling Wrch1 to cellular signaling, we analyzed in this study nWrch1 interaction with Src homology 3 (SH3) domains of different adaptor proteins. Using sedimentation and isothermal titration calorimetric (ITC) measurements, we identified isolated SH3 domains of growth factor receptor-bound protein 2 (Grb2), noncatalytic region of tyrosine kinase adaptor protein 1 (Nck1), c-Src, chicken tumor virus no. 10 (CT 10) regulator kinase 1 (Crk1), and p120 as low-affinity Wrch1-binding partners. Interestingly, under cell-based conditions, nWrch1 bound tightly to endogenous Grb2 and Nck, but not to Crk, c-Src, or p120. Consistent with this, a very tight nWrch1 interaction with full-length Grb2 and Nck1 was confirmed in vitro by ITC measurements indicating that high avidity of the adaptor proteins can compensate for the low affinity of their SH3 domains. Peptide analysis revealed that the central PxxP motif of nWrch1, which employs a minimal consensus sequence of eight amino acids with an essential arginine next to the PxxP motif, is responsible for these interactions. Thus, novel functional insights from this study suggest that multiple upstream signals may converge on Wrch1 directly through its SH3 domain-binding properties.
©2013 by Walter de Gruyter Berlin Boston
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Articles in the same Issue
- Masthead
- Masthead
- Guest Editorial
- Highlight: Kinin 2012 in Paris
- Highlight: Kinin 2012 in Paris
- Plasma kallikrein-kinin system and diabetic retinopathy
- Tissue kallikrein, blood pressure regulation, and hypertension: insight from genetic kallikrein deficiency
- Carboxypeptidase M augments kinin B1 receptor signaling by conformational crosstalk and enhances endothelial nitric oxide output
- Interactions between BdkrB2 and p53 genes in the developing kidney
- Further pharmacological evaluation of a novel synthetic peptide bradykinin B2 receptor agonist
- Role of kinin B2 receptors in opioid-induced hyperalgesia in inflammatory pain in mice
- Clinical impact of an angiotensin I-converting enzyme insertion/deletion and kinin B2 receptor +9/-9 polymorphisms in the prognosis of renal transplantation
- Bifunctional epitope-agonist ligands of the bradykinin B2 receptor
- Pro-angiogenic effect of human kallikrein-related peptidase 12 (KLK12) in lung endothelial cells does not depend on kinin-mediated activation of B2 receptor
- Review
- Mitochondrial pathways in sarcopenia of aging and disuse muscle atrophy
- Research Articles/Short Communications
- Genes and Nucleic Acids
- A genetic variant in the promoter region of miR-34b/c is associated with a reduced risk of colorectal cancer
- Protein Structure and Function
- SH3-mediated targeting of Wrch1/RhoU by multiple adaptor proteins
