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Title: A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis

Abstract

The gene encoding a putative siderophore-interacting protein from the marine bacterium S. frigidimarina was successfully cloned, followed by expression and purification of the gene product. Optimized crystals diffracted to 1.35 Å resolution and preliminary crystallographic analysis is promising with respect to structure determination and increased insight into the poorly understood molecular mechanisms underlying iron acquisition. Siderophore-binding proteins (SIPs) perform a key role in iron acquisition in multiple organisms. In the genome of the marine bacterium Shewanella frigidimarina NCIMB 400, the gene tagged as SFRI-RS12295 encodes a protein from this family. Here, the cloning, expression, purification and crystallization of this protein are reported, together with its preliminary X-ray crystallographic analysis to 1.35 Å resolution. The SIP crystals belonged to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 48.04, b = 78.31, c = 67.71 Å, α = 90, β = 99.94, γ = 90°, and are predicted to contain two molecules per asymmetric unit. Structure determination by molecular replacement and the use of previously determined ∼2 Å resolution SIP structures with ∼30% sequence identity as templates are ongoing.

Authors:
;  [1];  [1];  [2]; ;  [1]
  1. Universidade Nova de Lisboa, Avenida da República (EAN), 2780-157 Oeiras (Portugal)
  2. (iBET), Apartado 12, 2780-901 Oeiras (Portugal)
Publication Date:
OSTI Identifier:
22531141
Resource Type:
Journal Article
Resource Relation:
Journal Name: Acta Crystallographica. Section F, Structural Biology Communications; Journal Volume: 72; Journal Issue: Pt 9; Other Information: PMCID: PMC5012204; PMID: 27599855; PUBLISHER-ID: wd5266; PUBLISHER-ID: S2053230X16011419; OAI: oai:pubmedcentral.nih.gov:5012204; Copyright (c) Trindade et al. 2016; This is an open-access article distributed under the terms of the Creative Commons Attribution Licence, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are cited.; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTALLIZATION; CRYSTALLOGRAPHY; MONOCLINIC LATTICES; PROTEINS; RESOLUTION; SPACE GROUPS; X RADIATION; X-RAY DIFFRACTION

Citation Formats

Trindade, Inês B., Fonseca, Bruno M., Matias, Pedro M., Instituto de Biologia Experimental e Tecnológica, Louro, Ricardo O., and Moe, Elin, E-mail: elinmoe@itqb.unl.pt. A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis. United States: N. p., 2016. Web. doi:10.1107/S2053230X16011419.
Trindade, Inês B., Fonseca, Bruno M., Matias, Pedro M., Instituto de Biologia Experimental e Tecnológica, Louro, Ricardo O., & Moe, Elin, E-mail: elinmoe@itqb.unl.pt. A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis. United States. doi:10.1107/S2053230X16011419.
Trindade, Inês B., Fonseca, Bruno M., Matias, Pedro M., Instituto de Biologia Experimental e Tecnológica, Louro, Ricardo O., and Moe, Elin, E-mail: elinmoe@itqb.unl.pt. Tue . "A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis". United States. doi:10.1107/S2053230X16011419.
@article{osti_22531141,
title = {A putative siderophore-interacting protein from the marine bacterium Shewanella frigidimarina NCIMB 400: cloning, expression, purification, crystallization and X-ray diffraction analysis},
author = {Trindade, Inês B. and Fonseca, Bruno M. and Matias, Pedro M. and Instituto de Biologia Experimental e Tecnológica and Louro, Ricardo O. and Moe, Elin, E-mail: elinmoe@itqb.unl.pt},
abstractNote = {The gene encoding a putative siderophore-interacting protein from the marine bacterium S. frigidimarina was successfully cloned, followed by expression and purification of the gene product. Optimized crystals diffracted to 1.35 Å resolution and preliminary crystallographic analysis is promising with respect to structure determination and increased insight into the poorly understood molecular mechanisms underlying iron acquisition. Siderophore-binding proteins (SIPs) perform a key role in iron acquisition in multiple organisms. In the genome of the marine bacterium Shewanella frigidimarina NCIMB 400, the gene tagged as SFRI-RS12295 encodes a protein from this family. Here, the cloning, expression, purification and crystallization of this protein are reported, together with its preliminary X-ray crystallographic analysis to 1.35 Å resolution. The SIP crystals belonged to the monoclinic space group P2{sub 1}, with unit-cell parameters a = 48.04, b = 78.31, c = 67.71 Å, α = 90, β = 99.94, γ = 90°, and are predicted to contain two molecules per asymmetric unit. Structure determination by molecular replacement and the use of previously determined ∼2 Å resolution SIP structures with ∼30% sequence identity as templates are ongoing.},
doi = {10.1107/S2053230X16011419},
journal = {Acta Crystallographica. Section F, Structural Biology Communications},
number = Pt 9,
volume = 72,
place = {United States},
year = {Tue Aug 09 00:00:00 EDT 2016},
month = {Tue Aug 09 00:00:00 EDT 2016}
}