9AYC
Tetra-phosphorylated, E1435Q Ycf1 mutant in inward-facing wide conformation
Summary for 9AYC
| Entry DOI | 10.2210/pdb9ayc/pdb |
| EMDB information | 43985 |
| Descriptor | Metal resistance protein YCF1 (1 entity in total) |
| Functional Keywords | abc-transporter, ycf1, r-domain, phosphorylation, heavy metal, membrane protein |
| Biological source | Saccharomyces cerevisiae (brewer's yeast) |
| Total number of polymer chains | 1 |
| Total formula weight | 176263.23 |
| Authors | Carvalho, R.S.A.,Rasel, M.S.I.,Khandelwal, N.K.,Tomasiak, T.M. (deposition date: 2024-03-07, release date: 2024-08-21, Last modification date: 2025-09-03) |
| Primary citation | Souza Amado de Carvalho, R.,Rasel, M.S.I.,Khandelwal, N.K.,Tomasiak, T.M. Cryo-EM reveals a phosphorylated R-domain envelops the NBD1 catalytic domain in an ABC transporter. Life Sci Alliance, 7:-, 2024 Cited by PubMed Abstract: Many ATP-binding cassette transporters are regulated by phosphorylation on long and disordered loops which presents a challenge to visualize with structural methods. We have trapped an activated state of the regulatory domain (R-domain) of yeast cadmium factor 1 (Ycf1) by enzymatically enriching the phosphorylated state. A 3.23 Å cryo-EM structure reveals an R-domain structure with four phosphorylated residues and the position for the entire R-domain. The structure reveals key R-domain interactions including a bridging interaction between NBD1 and NBD2 and an interaction with the R-insertion, another regulatory region. We scanned these interactions by systematically replacing segments along the entire R-domain with scrambled combinations of alanine, glycine, and glutamine and probing function under cellular conditions that require the Ycf1 function. We find a close match with these interactions and interacting regions on our R-domain structure that points to the importance of most well-structured segments for function. We propose a model where the R-domain stabilizes a transport-competent state upon phosphorylation by enveloping NBD1 entirely. PubMed: 39209537DOI: 10.26508/lsa.202402779 PDB entries with the same primary citation |
| Experimental method | ELECTRON MICROSCOPY (3.23 Å) |
Structure validation
Download full validation report
