High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane.

James D. Zook, Trivikram R. Molugu, Neil E. Jacobsen, Guangxin Lin, Jürgen Soll, Brian R. Cherry, Michael F. Brown, Petra Fromme

Research output: Contribution to journalArticle

Abstract

Solving high-resolution structures for membrane proteins continues to be a daunting challenge in the structural biology community. In this study we report our high-resolution NMR results for a transmembrane protein, outer envelope protein of molar mass 16 kDa (OEP16), an amino acid transporter from the outer membrane of chloroplasts. Three-dimensional, high-resolution NMR experiments on the (13)C, (15)N, (2)H-triply-labeled protein were used to assign protein backbone resonances and to obtain secondary structure information. The results yield over 95% assignment of N, HN, CO, Cα, and Cβ chemical shifts, which is essential for obtaining a high resolution structure from NMR data. Chemical shift analysis from the assignment data reveals experimental evidence for the first time on the location of the secondary structure elements on a per residue basis. In addition T 1Z and T2 relaxation experiments were performed in order to better understand the protein dynamics. Arginine titration experiments yield an insight into the amino acid residues responsible for protein transporter function. The results provide the necessary basis for high-resolution structural determination of this important plant membrane protein.

Original languageEnglish
Article numbere78116
JournalPLoS One
Volume8
Issue number10
Publication statusPublished - 2013

Fingerprint

amino acid transporters
Amino Acid Transport Systems
Chloroplasts
chloroplasts
Nuclear magnetic resonance
Membranes
Proteins
proteins
membrane proteins
Chemical shift
Membrane Proteins
transmembrane proteins
plant proteins
Plant Proteins
titration
Molar mass
Experiments
arginine
transporters
Carbon Monoxide

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Zook, J. D., Molugu, T. R., Jacobsen, N. E., Lin, G., Soll, J., Cherry, B. R., ... Fromme, P. (2013). High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane. PLoS One, 8(10), [e78116].

High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane. / Zook, James D.; Molugu, Trivikram R.; Jacobsen, Neil E.; Lin, Guangxin; Soll, Jürgen; Cherry, Brian R.; Brown, Michael F.; Fromme, Petra.

In: PLoS One, Vol. 8, No. 10, e78116, 2013.

Research output: Contribution to journalArticle

Zook, JD, Molugu, TR, Jacobsen, NE, Lin, G, Soll, J, Cherry, BR, Brown, MF & Fromme, P 2013, 'High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane.', PLoS One, vol. 8, no. 10, e78116.
Zook JD, Molugu TR, Jacobsen NE, Lin G, Soll J, Cherry BR et al. High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane. PLoS One. 2013;8(10). e78116.
Zook, James D. ; Molugu, Trivikram R. ; Jacobsen, Neil E. ; Lin, Guangxin ; Soll, Jürgen ; Cherry, Brian R. ; Brown, Michael F. ; Fromme, Petra. / High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane. In: PLoS One. 2013 ; Vol. 8, No. 10.
@article{7479b73b840a4214808c7a2d6dfc834c,
title = "High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane.",
abstract = "Solving high-resolution structures for membrane proteins continues to be a daunting challenge in the structural biology community. In this study we report our high-resolution NMR results for a transmembrane protein, outer envelope protein of molar mass 16 kDa (OEP16), an amino acid transporter from the outer membrane of chloroplasts. Three-dimensional, high-resolution NMR experiments on the (13)C, (15)N, (2)H-triply-labeled protein were used to assign protein backbone resonances and to obtain secondary structure information. The results yield over 95{\%} assignment of N, HN, CO, Cα, and Cβ chemical shifts, which is essential for obtaining a high resolution structure from NMR data. Chemical shift analysis from the assignment data reveals experimental evidence for the first time on the location of the secondary structure elements on a per residue basis. In addition T 1Z and T2 relaxation experiments were performed in order to better understand the protein dynamics. Arginine titration experiments yield an insight into the amino acid residues responsible for protein transporter function. The results provide the necessary basis for high-resolution structural determination of this important plant membrane protein.",
author = "Zook, {James D.} and Molugu, {Trivikram R.} and Jacobsen, {Neil E.} and Guangxin Lin and J{\"u}rgen Soll and Cherry, {Brian R.} and Brown, {Michael F.} and Petra Fromme",
year = "2013",
language = "English",
volume = "8",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "10",

}

TY - JOUR

T1 - High-resolution NMR reveals secondary structure and folding of amino acid transporter from outer chloroplast membrane.

AU - Zook, James D.

AU - Molugu, Trivikram R.

AU - Jacobsen, Neil E.

AU - Lin, Guangxin

AU - Soll, Jürgen

AU - Cherry, Brian R.

AU - Brown, Michael F.

AU - Fromme, Petra

PY - 2013

Y1 - 2013

N2 - Solving high-resolution structures for membrane proteins continues to be a daunting challenge in the structural biology community. In this study we report our high-resolution NMR results for a transmembrane protein, outer envelope protein of molar mass 16 kDa (OEP16), an amino acid transporter from the outer membrane of chloroplasts. Three-dimensional, high-resolution NMR experiments on the (13)C, (15)N, (2)H-triply-labeled protein were used to assign protein backbone resonances and to obtain secondary structure information. The results yield over 95% assignment of N, HN, CO, Cα, and Cβ chemical shifts, which is essential for obtaining a high resolution structure from NMR data. Chemical shift analysis from the assignment data reveals experimental evidence for the first time on the location of the secondary structure elements on a per residue basis. In addition T 1Z and T2 relaxation experiments were performed in order to better understand the protein dynamics. Arginine titration experiments yield an insight into the amino acid residues responsible for protein transporter function. The results provide the necessary basis for high-resolution structural determination of this important plant membrane protein.

AB - Solving high-resolution structures for membrane proteins continues to be a daunting challenge in the structural biology community. In this study we report our high-resolution NMR results for a transmembrane protein, outer envelope protein of molar mass 16 kDa (OEP16), an amino acid transporter from the outer membrane of chloroplasts. Three-dimensional, high-resolution NMR experiments on the (13)C, (15)N, (2)H-triply-labeled protein were used to assign protein backbone resonances and to obtain secondary structure information. The results yield over 95% assignment of N, HN, CO, Cα, and Cβ chemical shifts, which is essential for obtaining a high resolution structure from NMR data. Chemical shift analysis from the assignment data reveals experimental evidence for the first time on the location of the secondary structure elements on a per residue basis. In addition T 1Z and T2 relaxation experiments were performed in order to better understand the protein dynamics. Arginine titration experiments yield an insight into the amino acid residues responsible for protein transporter function. The results provide the necessary basis for high-resolution structural determination of this important plant membrane protein.

UR - http://www.scopus.com/inward/record.url?scp=84907042040&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84907042040&partnerID=8YFLogxK

M3 - Article

VL - 8

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 10

M1 - e78116

ER -