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

Standard

Structure determination of a major facilitator peptide transporter : Inward facing PepTSt from Streptococcus thermophilus crystallized in space group P3121. / Quistgaard, Esben M; Martinez Molledo, Maria; Löw, Christian.

In: P L o S One, Vol. 12, No. 3, 06.03.2017, p. e0173126.

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

Harvard

APA

CBE

MLA

Vancouver

Author

Bibtex

@article{8e409e011aa2488db7332b5180a57c83,
title = "Structure determination of a major facilitator peptide transporter: Inward facing PepTSt from Streptococcus thermophilus crystallized in space group P3121",
abstract = "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.",
keywords = "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",
author = "Quistgaard, {Esben M} and {Martinez Molledo}, Maria and Christian L{\"o}w",
year = "2017",
month = "3",
day = "6",
doi = "10.1371/journal.pone.0173126",
language = "English",
volume = "12",
pages = "e0173126",
journal = "P L o S One",
issn = "1932-6203",
publisher = "public library of science",
number = "3",

}

RIS

TY - JOUR

T1 - Structure determination of a major facilitator peptide transporter

T2 - Inward facing PepTSt from Streptococcus thermophilus crystallized in space group P3121

AU - Quistgaard, Esben M

AU - Martinez Molledo, Maria

AU - Löw, Christian

PY - 2017/3/6

Y1 - 2017/3/6

N2 - 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.

AB - 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.

KW - Amino Acid Motifs

KW - Amino Acid Sequence

KW - Bacterial Proteins

KW - Membrane Transport Proteins

KW - Models, Molecular

KW - Peptides

KW - Protein Conformation

KW - Protein Multimerization

KW - Protein Stability

KW - Streptococcus thermophilus

KW - Structure-Activity Relationship

KW - Thermodynamics

KW - Journal Article

U2 - 10.1371/journal.pone.0173126

DO - 10.1371/journal.pone.0173126

M3 - Journal article

C2 - 28264013

VL - 12

SP - e0173126

JO - P L o S One

JF - P L o S One

SN - 1932-6203

IS - 3

ER -