PDBsum entry 5d3m

Transport protein

PDB id: 5d3m

Contents

Protein chains

280 a.a.

281 a.a.

168 a.a.

259 a.a.

Ligands

ANP ×4

PDB id:

5d3m

Name:

Transport protein

Title:

Folate ecf transporter: amppnp bound state

Structure:

Energy-coupling factor transporter atp-binding protein ecfa1. Chain: a, e. Synonym: ecf transporter a component ecfa1. Engineered: yes. Other_details: 10his-tag with tev cleavage site at n-terminus. Energy-coupling factor transporter atp-binding protein ecfa2. Chain: b, f.

Source:

Lactobacillus delbrueckii. Organism_taxid: 1584. Gene: ecfa1, cbio1, ldb0424. Expressed in: escherichia coli. Expression_system_taxid: 562. Gene: ecfa2, cbio2, ldb0425. Gene: ldb1625. Gene: cbiq, ecft, lbvib27_01965, lbvib44_01960, sb57_06620. Expression_system_taxid: 562

Resolution:

3.30Å R-factor: 0.252 R-free: 0.289

Authors:

A.Guskov,L.J.Y.M.Swier,D.J.Slotboom

Key ref:

L.J.Swier et al. (2016). Structural insight in the toppling mechanism of an energy-coupling factor transporter. Nat Commun, 7, 11072. PubMed id: 27026363 DOI: 10.1038/ncomms11072

Date:

06-Aug-15 Release date: 06-Apr-16

Protein chains

Q1GBJ0  (ECFA1_LACDA) -  Energy-coupling factor transporter ATP-binding protein EcfA1 from Lactobacillus delbrueckii subsp. bulgaricus (strain ATCC 11842 / DSM 20081 / BCRC 10696 / JCM 1002 / NBRC 13953 / NCIMB 11778 / NCTC 12712 / WDCM 00102 / Lb 14)

Seq: 282 a.a.

Struc: 280 a.a.*

Protein chains

Q1GBI9  (ECFA2_LACDA) -  Energy-coupling factor transporter ATP-binding protein EcfA2 from Lactobacillus delbrueckii subsp. bulgaricus (strain ATCC 11842 / DSM 20081 / BCRC 10696 / JCM 1002 / NBRC 13953 / NCIMB 11778 / NCTC 12712 / WDCM 00102 / Lb 14)

Seq: 287 a.a.

Struc: 281 a.a.

Protein chains

Q1G929  (Q1G929_LACDA) -  Conserved hypothetical membrane protein from Lactobacillus delbrueckii subsp. bulgaricus (strain ATCC 11842 / DSM 20081 / BCRC 10696 / JCM 1002 / NBRC 13953 / NCIMB 11778 / NCTC 12712 / WDCM 00102 / Lb 14)

Seq: 176 a.a.

Struc: 168 a.a.

Protein chains

Q1GBI8  (Q1GBI8_LACDA) -  Energy-coupling factor transporter transmembrane protein EcfT from Lactobacillus delbrueckii subsp. bulgaricus (strain ATCC 11842 / DSM 20081 / BCRC 10696 / JCM 1002 / NBRC 13953 / NCIMB 11778 / NCTC 12712 / WDCM 00102 / Lb 14)

Seq: 265 a.a.

Struc: 259 a.a.

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: Chains A, B, E, F: E.C.7.-.-.-

DOI no: 10.1038/ncomms11072

Nat Commun 7:11072 (2016)

PubMed id: 27026363

Structural insight in the toppling mechanism of an energy-coupling factor transporter.

L.J.Swier, A.Guskov, D.J.Slotboom.

ABSTRACT

Energy-coupling factor (ECF) transporters mediate uptake of micronutrients in prokaryotes. The transporters consist of an S-component that binds the transported substrate and an ECF module (EcfAA'T) that binds and hydrolyses ATP. The mechanism of transport is poorly understood but presumably involves an unusual step in which the membrane-embedded S-component topples over to carry the substrate across the membrane. In many ECF transporters, the S-component dissociates from the ECF module after transport. Subsequently, substrate-bound S-components out-compete the empty proteins for re-binding to the ECF module in a new round of transport. Here we present crystal structures of the folate-specific transporter ECF-FolT from Lactobacillus delbrueckii. Interaction of the ECF module with FolT stabilizes the toppled state, and simultaneously destroys the high-affinity folate-binding site, allowing substrate release into the cytosol. We hypothesize that differences in the kinetics of toppling can explain how substrate-loaded FolT out-competes apo-FolT for association with the ECF module.