4XH9
CRYSTAL STRUCTURE OF HUMAN RHOA IN COMPLEX WITH DH/PH FRAGMENT OF THE GUANINE NUCLEOTIDE EXCHANGE FACTOR NET1
Summary for 4XH9
| Entry DOI | 10.2210/pdb4xh9/pdb |
| Descriptor | Neuroepithelial cell-transforming gene 1 protein, Transforming protein RhoA (3 entities in total) |
| Functional Keywords | rhoa gtpase, signaling protein, activator, guanine nucleotide exchange factor |
| Biological source | Homo sapiens (Human) More |
| Total number of polymer chains | 4 |
| Total formula weight | 125043.09 |
| Authors | Garcia, C.,Petit, P.,Boutin, J.A.,Ferry, G.,Vuillard, L. (deposition date: 2015-01-05, release date: 2015-01-14, Last modification date: 2024-01-10) |
| Primary citation | Petit, A.P.,Garcia-Petit, C.,Bueren-Calabuig, J.A.,Vuillard, L.M.,Ferry, G.,Boutin, J.A. A structural study of the complex between neuroepithelial cell transforming gene 1 (Net1) and RhoA reveals a potential anticancer drug hot spot. J. Biol. Chem., 293:9064-9077, 2018 Cited by PubMed Abstract: The GTPase RhoA is a major player in many different regulatory pathways. RhoA catalyzes GTP hydrolysis, and its catalysis is accelerated when RhoA forms heterodimers with proteins of the guanine nucleotide exchange factor (GEF) family. Neuroepithelial cell transforming gene 1 (Net1) is a RhoA-interacting GEF implicated in cancer, but the structural features supporting the RhoA/Net1 interaction are unknown. Taking advantage of a simple production and purification process, here we solved the structure of a RhoA/Net1 heterodimer with X-ray crystallography at 2-Å resolution. Using a panel of several techniques, including molecular dynamics simulations, we characterized the RhoA/Net1 interface. Moreover, deploying an extremely simple peptide-based scanning approach, we found that short peptides (penta- to nonapeptides) derived from the protein/protein interaction region of RhoA could disrupt the RhoA/Net1 interaction and thereby diminish the rate of nucleotide exchange. The most inhibitory peptide, EVKHF, spanning residues 102-106 in the RhoA sequence, displayed an IC of ∼100 μm without further modifications. The peptides identified here could be useful in further investigations of the RhoA/Net1 interaction region. We propose that our structural and functional insights might inform chemical approaches for transforming the pentapeptide into an optimized pseudopeptide that antagonizes Net1-mediated RhoA activation with therapeutic anticancer potential. PubMed: 29695506DOI: 10.1074/jbc.RA117.001123 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2 Å) |
Structure validation
Download full validation report
