Journal of Biomolecular NMR

, Volume 7, Issue 3, pp 225–235

1H and 15N NMR resonance assignments and solution secondary structure of oxidized Desulfovibrio desulfuricans flavodoxin

  • John R. Pollock
  • Richard P. Swenson
  • Brian J. Stockman
Research Papers

Summary

Sequence-specific 1H and 15N resonance assignments have been made for 137 of the 146 nonprolyl residues in oxidized Desulfovibrio desulfuricans [Essex 6] flavodoxin. Assignments were obtained by a concerted analysis of the heteronuclear three-dimensional 1H-15N NOESY-HMQC and TOCSY-HMQC data sets, recorded on uniformly 15N-enriched protein at 300 K. Numerous side-chain resonances have been partially or fully assigned. Residues with overlapping 1HN chemical shifts were resolved by a three-dimensional 1H-15N HMQC-NOESY-HMQC spectrum. Medium-and long-range NOEs, 3JNHα coupling constants, and 1HN exchange data indicate a secondary structure consisting of five parallel β-strands and four α-helices with a topology similar to that of Desulfovibrio vulgaris [Hidenborough] flavodoxin. Prolines at positions 106 and 134, which are not conserved in D. vulgaris flavodoxin, contort the two C-terminal α-helices.

Keywords

Chemical shift index Flavodoxin Isotopic enrichment Nitrogen-15 Protein Secondary structure 3D NMR 

Abbreviations

CSI

chemical shift index

DQF-COSY

double-quantum-filtered correlation spectroscopy

DIPSI

decoupling in the presence of scalar interactions

FMN

flavin mononucleotide

GARP

globally optimized alternating phase rectangular pulse

HMQC

heteronuclear multiple-quantum coherence

HSQC

heteronuclear single-quantum coherence

NOE

nuclear Overhauser effect

NOESY

nuclear Overhauser enhancement spectroscopy

TOCSY

total correlation spectroscopy

TPPI

time-proportional phase increments

TSP

3-(trimethylsilyl)propionic-2,2,3,3-d4 acid, sodium salt

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Copyright information

© ESCOM Science Publishers B.V 1996

Authors and Affiliations

  • John R. Pollock
    • 1
  • Richard P. Swenson
    • 2
  • Brian J. Stockman
    • 1
  1. 1.Pharmacia and Upjohn, Inc.KalamazooU.S.A.
  2. 2.Department of BiochemistryThe Ohio State UniversityColumbusU.S.A.

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