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Title: Structure Determination of an FMN Reductase from Pseudomonas aeruginosa PA01 using Sulfur Anomalous Signal

Abstract

The availability of high-intensity synchrotron facilities, technological advances in data-collection techniques and improved data-reduction and crystallographic software have ushered in a new era in high-throughput macromolecular crystallography. Here, the de novo automated crystal structure determination at 1.28 Angstroms resolution of an NAD(P)H-dependent FMN reductase flavoprotein from Pseudomonas aeruginosa PA01-derived protein Q9I4D4 using the anomalous signal from an unusually small number of S atoms is reported. Although this protein lacks the flavodoxin key fingerprint motif [(T/S)XTGXT], it has been confirmed to bind flavin mononucleotide and the binding site was identified via X-ray crystallography. This protein contains a novel flavin mononucleotide-binding site GSLRSGSYN, which has not been previously reported. Detailed statistics pertaining to sulfur phasing and other factors contributing to structure determination are discussed. Structural comparisons of the apoenzyme and the protein complexed with flavin mononucleotide show conformational changes on cofactor binding. NADPH-dependent activity has been confirmed with biochemical assays.

Authors:
; ; ;
Publication Date:
Research Org.:
Brookhaven National Laboratory (BNL) National Synchrotron Light Source
Sponsoring Org.:
Doe - Office Of Science
OSTI Identifier:
914143
Report Number(s):
BNL-78711-2007-JA
TRN: US0801572
DOE Contract Number:  
DE-AC02-98CH10886
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Cryst. D; Journal Volume: 62
Country of Publication:
United States
Language:
English
Subject:
36 MATERIALS SCIENCE; 43 PARTICLE ACCELERATORS; ATOMS; AVAILABILITY; CONFORMATIONAL CHANGES; CRYSTAL STRUCTURE; CRYSTALLOGRAPHY; ISOALLOXAZINES; OXIDOREDUCTASES; PROTEINS; PSEUDOMONAS; RESOLUTION; STATISTICS; SULFUR; SYNCHROTRONS; national synchrotron light source

Citation Formats

Agarwal,R., Bonanno, J., Burley, S., and Swaminathan, S. Structure Determination of an FMN Reductase from Pseudomonas aeruginosa PA01 using Sulfur Anomalous Signal. United States: N. p., 2006. Web. doi:10.1107/S0907444906001600.
Agarwal,R., Bonanno, J., Burley, S., & Swaminathan, S. Structure Determination of an FMN Reductase from Pseudomonas aeruginosa PA01 using Sulfur Anomalous Signal. United States. doi:10.1107/S0907444906001600.
Agarwal,R., Bonanno, J., Burley, S., and Swaminathan, S. Sun . "Structure Determination of an FMN Reductase from Pseudomonas aeruginosa PA01 using Sulfur Anomalous Signal". United States. doi:10.1107/S0907444906001600.
@article{osti_914143,
title = {Structure Determination of an FMN Reductase from Pseudomonas aeruginosa PA01 using Sulfur Anomalous Signal},
author = {Agarwal,R. and Bonanno, J. and Burley, S. and Swaminathan, S.},
abstractNote = {The availability of high-intensity synchrotron facilities, technological advances in data-collection techniques and improved data-reduction and crystallographic software have ushered in a new era in high-throughput macromolecular crystallography. Here, the de novo automated crystal structure determination at 1.28 Angstroms resolution of an NAD(P)H-dependent FMN reductase flavoprotein from Pseudomonas aeruginosa PA01-derived protein Q9I4D4 using the anomalous signal from an unusually small number of S atoms is reported. Although this protein lacks the flavodoxin key fingerprint motif [(T/S)XTGXT], it has been confirmed to bind flavin mononucleotide and the binding site was identified via X-ray crystallography. This protein contains a novel flavin mononucleotide-binding site GSLRSGSYN, which has not been previously reported. Detailed statistics pertaining to sulfur phasing and other factors contributing to structure determination are discussed. Structural comparisons of the apoenzyme and the protein complexed with flavin mononucleotide show conformational changes on cofactor binding. NADPH-dependent activity has been confirmed with biochemical assays.},
doi = {10.1107/S0907444906001600},
journal = {Acta Cryst. D},
number = ,
volume = 62,
place = {United States},
year = {Sun Jan 01 00:00:00 EST 2006},
month = {Sun Jan 01 00:00:00 EST 2006}
}