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

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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, 3JNH α 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.

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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-d 4 acid, sodium salt

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Author notes

  1. Brian J. Stockman

    Present address: Pharmacia and Upjohn, Inc., 301 Henrietta Street, 49001-0199, Kalamazoo, MI, U.S.A.

Authors and Affiliations

  1. Pharmacia and Upjohn, Inc., 301 Henrietta Street, 49001-0199, Kalamazoo, MI, U.S.A.

    John R. Pollock

  2. Department of Biochemistry, The Ohio State University, 43210, Columbus, OH, U.S.A.

    Richard P. Swenson

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  1. John R. Pollock
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  2. Richard P. Swenson
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Pollock, J.R., Swenson, R.P. & Stockman, B.J. 1H and 15N NMR resonance assignments and solution secondary structure of oxidized Desulfovibrio desulfuricans flavodoxin. J Biomol NMR 7, 225–235 (1996). https://doi.org/10.1007/BF00202039

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  • DOI: https://doi.org/10.1007/BF00202039

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