7M69

E1435Q Ycf1 mutant in inward-facing wide conformation

Summary for 7M69

• Entry DOI: 10.2210/pdb7m69/pdb
• EMDB information: 23691
• Descriptor: Metal resistance protein YCF1 (1 entity in total)
• Functional Keywords: abc transporter, membrane protein
• Biological source: Saccharomyces cerevisiae S288C
• Total number of polymer chains: 1
• Total formula weight: 176318.33

Authors

Khandelwal, N.K.,Millan, C.R.,Thaker, T.M.,Tomasiak, T.M. (deposition date: 2021-03-25, release date: 2022-04-06, Last modification date: 2024-10-16)

Primary citation

Khandelwal, N.K.,Millan, C.R.,Zangari, S.I.,Avila, S.,Williams, D.,Thaker, T.M.,Tomasiak, T.M.
The structural basis for regulation of the glutathione transporter Ycf1 by regulatory domain phosphorylation.
Nat Commun, 13:1278-1278, 2022

PubMed Abstract: Yeast Cadmium Factor 1 (Ycf1) sequesters heavy metals and glutathione into the vacuole to counter cell stress. Ycf1 belongs to the ATP binding cassette C-subfamily (ABCC) of transporters, many of which are regulated by phosphorylation on intrinsically-disordered domains. The regulatory mechanism of phosphorylation is still poorly understood. Here, we report two cryo-EM structures of Ycf1 at 3.4 Å and 4.0 Å resolution in inward-facing open conformations that capture previously unobserved ordered states of the intrinsically disordered regulatory domain (R-domain). R-domain phosphorylation is clearly evident and induces a topology promoting electrostatic and hydrophobic interactions with Nucleotide Binding Domain 1 (NBD1) and the Lasso motif. These interactions stay constant between the structures and are related by rigid body movements of the NBD1/R-domain complex. Biochemical data further show R-domain phosphorylation reorganizes the Ycf1 architecture and is required for maximal ATPase activity. Together, we provide insights into how R-domains control ABCC transporter activity.

PubMed: 35277487
DOI: 10.1038/s41467-022-28811-w

Experimental method

ELECTRON MICROSCOPY (3.42 Å)