Aarhus University Seal / Aarhus Universitets segl

Esben Meldgaard Høgh Quistgaard

Structure determination of a major facilitator peptide transporter: Inward facing PepTSt from Streptococcus thermophilus crystallized in space group P3121

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

  • Esben M Quistgaard
  • Maria Martinez Molledo, Centre for Structural Systems Biology (CSSB), DESY and European Molecular Biology Laboratory Hamburg, Hamburg, Germany.
  • ,
  • Christian Löw, Department of Medical Biochemistry and Biophysics, Division of Physiological Chemistry II, Karolinska Institutet, Stockholm, Sweden.

Major facilitator superfamily (MFS) peptide transporters (typically referred to as PepT, POT or PTR transporters) mediate the uptake of di- and tripeptides, and so play an important dietary role in many organisms. In recent years, a better understanding of the molecular basis for this process has emerged, which is in large part due to a steep increase in structural information. Yet, the conformational transitions underlying the transport mechanism are still not fully understood, and additional data is therefore needed. Here we report in detail the detergent screening, crystallization, experimental MIRAS phasing, and refinement of the peptide transporter PepTSt from Streptococcus thermophilus. The space group is P3121, and the protein is crystallized in a monomeric inward facing form. The binding site is likely to be somewhat occluded, as the lobe encompassing transmembrane helices 10 and 11 is markedly bent towards the central pore of the protein, but the extent of this potential occlusion could not be determined due to disorder at the apex of the lobe. Based on structural comparisons with the seven previously determined P212121 and C2221 structures of inward facing PepTSt, the structural flexibility as well as the conformational changes mediating transition between the inward open and inward facing occluded states are discussed. In conclusion, this report improves our understanding of the structure and conformational cycle of PepTSt, and can furthermore serve as a case study, which may aid in supporting future structure determinations of additional MFS transporters or other integral membrane proteins.

Original languageEnglish
JournalP L o S One
Volume12
Issue3
Pages (from-to)e0173126
ISSN1932-6203
DOIs
Publication statusPublished - 6 Mar 2017
Externally publishedYes

    Research areas

  • Amino Acid Motifs, Amino Acid Sequence, Bacterial Proteins, Membrane Transport Proteins, Models, Molecular, Peptides, Protein Conformation, Protein Multimerization, Protein Stability, Streptococcus thermophilus, Structure-Activity Relationship, Thermodynamics, Journal Article

See relations at Aarhus University Citationformats

ID: 118858597