PDBsum entry 4hul

Transport protein

PDB id: 4hul

Contents

Protein chains

459 a.a.

91 a.a.

Metals

_CS

PDB id:

4hul

Name:

Transport protein

Title:

Mate transporter norm-ng in complex with cs+ and monobody

Structure:

Multidrug efflux protein. Chain: a. Engineered: yes. Protein b. Chain: b. Engineered: yes

Source:

Neisseria gonorrhoeae. Organism_taxid: 940296. Gene: ngtw08_0430. Expressed in: escherichia coli. Expression_system_taxid: 562. Escherichia coli. Organism_taxid: 562. Expression_system_taxid: 562

Resolution:

3.81Å R-factor: 0.314 R-free: 0.376

Authors:

M.Lu

Key ref:

M.Lu et al. (2013). Structures of a Na+-coupled, substrate-bound MATE multidrug transporter. Proc Natl Acad Sci U S A, 110, 2099-2104. PubMed id: 23341609

Date:

02-Nov-12 Release date: 06-Feb-13

Protein chain

No UniProt id for this chain

Struc: 459 a.a.

Protein chain

M1E1G6  (M1E1G6_ECOLX) -  Protein B from Escherichia coli

Seq: 99 a.a.

Struc: 91 a.a.

Proc Natl Acad Sci U S A 110:2099-2104 (2013)

PubMed id: 23341609

Structures of a Na+-coupled, substrate-bound MATE multidrug transporter.

M.Lu, J.Symersky, M.Radchenko, A.Koide, Y.Guo, R.Nie, S.Koide.

ABSTRACT

Multidrug transporters belonging to the multidrug and toxic compound extrusion (MATE) family expel dissimilar lipophilic and cationic drugs across cell membranes by dissipating a preexisting Na(+) or H(+) gradient. Despite its clinical relevance, the transport mechanism of MATE proteins remains poorly understood, largely owing to a lack of structural information on the substrate-bound transporter. Here we report crystal structures of a Na(+)-coupled MATE transporter NorM from Neisseria gonorrheae in complexes with three distinct translocation substrates (ethidium, rhodamine 6G, and tetraphenylphosphonium), as well as Cs(+) (a Na(+) congener), all captured in extracellular-facing and drug-bound states. The structures revealed a multidrug-binding cavity festooned with four negatively charged amino acids and surprisingly limited hydrophobic moieties, in stark contrast to the general belief that aromatic amino acids play a prominent role in multidrug recognition. Furthermore, we discovered an uncommon cation-π interaction in the Na(+)-binding site located outside the drug-binding cavity and validated the biological relevance of both the substrate- and cation-binding sites by conducting drug resistance and transport assays. Additionally, we uncovered potential rearrangement of at least two transmembrane helices upon Na(+)-induced drug export. Based on our structural and functional analyses, we suggest that Na(+) triggers multidrug extrusion by inducing protein conformational changes rather than by directly competing for the substrate-binding amino acids. This scenario is distinct from the canonical antiport mechanism, in which both substrate and counterion compete for a shared binding site in the transporter. Collectively, our findings provide an important step toward a detailed and mechanistic understanding of multidrug transport.