GFT projection NMR based resonance assignment of membrane proteins: Application to subunit c of E. coli F1F0 ATP synthase in LPPG micelles

Qi Zhang, Hanudatta S. Atreya, Douglas E. Kamen, Mark E. Girvin, Thomas Szyperski

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

G-matrix FT projection NMR spectroscopy was employed for resonance assignment of the 79-residue subunit c of the Escherichia coli F1F0 ATP synthase embedded in micelles formed by lyso palmitoyl phosphatidyl glycerol (LPPG). Five GFT NMR experiments, that is, (3,2)D HN NCO, L-(4,3)D HNN Cαβ Cα, L-(4,3)D HNN(CO)Cαβ Cα, (4,2)D HACA(CO) NHN and (4,3)D HCCH, were acquired along with simultaneous 3D 15N, 13Caliphatic, 13Caromatic- resolved [1H,1H]-NOESY with a total measurement time of ∼43 h. Data analysis resulted in sequence specific assignments for all routinely measured backbone and 13Cβ shifts, and for 97% of the side chain shifts. Moreover, the use of two G2FT NMR experiments, that is, (5,3)D HN{NCO} {CαβCα} and (5,3)D Cαβ Cα} {CON}HN, was explored to break the very high chemical shift degeneracy typically encountered for membrane proteins. It is shown that the 4D and 5D spectral information obtained rapidly from GFT and G2FT NMR experiments enables one to efficiently obtain (nearly) complete resonance assignments of membrane proteins.

Original languageEnglish (US)
Pages (from-to)157-163
Number of pages7
JournalJournal of Biomolecular NMR
Volume40
Issue number3
DOIs
StatePublished - Mar 2008

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Dilatation and Curettage
Phosphatidylglycerols
Micelles
Carbon Monoxide
Escherichia coli
Membrane Proteins
Nuclear magnetic resonance
Magnetic Resonance Spectroscopy
Experiments
Chemical shift
Time measurement
Nuclear magnetic resonance spectroscopy
F1F0-ATP synthase
palmitoyl glycerol
2,2',2''-trichlorotriethylamine

Keywords

  • FFATP synthase
  • GFT NMR
  • LPPG
  • Membrane protein
  • Projection NMR
  • Resonance assignment
  • Subunitc

ASJC Scopus subject areas

  • Spectroscopy
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

GFT projection NMR based resonance assignment of membrane proteins : Application to subunit c of E. coli F1F0 ATP synthase in LPPG micelles. / Zhang, Qi; Atreya, Hanudatta S.; Kamen, Douglas E.; Girvin, Mark E.; Szyperski, Thomas.

In: Journal of Biomolecular NMR, Vol. 40, No. 3, 03.2008, p. 157-163.

Research output: Contribution to journalArticle

Zhang, Qi ; Atreya, Hanudatta S. ; Kamen, Douglas E. ; Girvin, Mark E. ; Szyperski, Thomas. / GFT projection NMR based resonance assignment of membrane proteins : Application to subunit c of E. coli F1F0 ATP synthase in LPPG micelles. In: Journal of Biomolecular NMR. 2008 ; Vol. 40, No. 3. pp. 157-163.
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