6VJF
The P-Loop K to A mutation of C. therm Vps1 GTPase-BSE
Summary for 6VJF
| Entry DOI | 10.2210/pdb6vjf/pdb |
| Related | 6DEF |
| Descriptor | Putative sorting protein Vps1, PHOSPHOMETHYLPHOSPHONIC ACID GUANYLATE ESTER, MAGNESIUM ION, ... (4 entities in total) |
| Functional Keywords | gtpase, dynamin, dynamin superfamily, bse, ged, gcp, p-loop, hydrolase |
| Biological source | Chaetomium thermophilum (strain DSM 1495 / CBS 144.50 / IMI 039719) More |
| Total number of polymer chains | 4 |
| Total formula weight | 173957.94 |
| Authors | Tornabene, B.A.,Varlakhanova, N.V.,Chappie, J.S.,Ford, M.G.J. (deposition date: 2020-01-15, release date: 2020-02-19, Last modification date: 2023-10-11) |
| Primary citation | Tornabene, B.A.,Varlakhanova, N.V.,Hosford, C.J.,Chappie, J.S.,Ford, M.G.J. Structural and functional characterization of the dominant negative P-loop lysine mutation in the dynamin superfamily protein Vps1. Protein Sci., 29:1416-1428, 2020 Cited by PubMed Abstract: Dynamin-superfamily proteins (DSPs) are large self-assembling mechanochemical GTPases that harness GTP hydrolysis to drive membrane remodeling events needed for many cellular processes. Mutation to alanine of a fully conserved lysine within the P-loop of the DSP GTPase domain results in abrogation of GTPase activity. This mutant has been widely used in the context of several DSPs as a dominant-negative to impair DSP-dependent processes. However, the precise deficit of the P-loop K to A mutation remains an open question. Here, we use biophysical, biochemical and structural approaches to characterize this mutant in the context of the endosomal DSP Vps1. We show that the Vps1 P-loop K to A mutant binds nucleotide with an affinity similar to wild type but exhibits defects in the organization of the GTPase active site that explain the lack of hydrolysis. In cells, Vps1 and Dnm1 bearing the P-loop K to A mutation are defective in disassembly. These mutants become trapped in assemblies at the typical site of action of the DSP. This work provides mechanistic insight into the widely-used DSP P-loop K to A mutation and the basis of its dominant-negative effects in the cell. PubMed: 31981262DOI: 10.1002/pro.3830 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.472 Å) |
Structure validation
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