High-throughput screening of structural proteomics targets using NMR

We applied a high-throughput strategy for the screening of targets for structural proteomics of Xanthomonas axonopodis pv citri. This strategy is based on the rapid 1H– 15N HSQC NMR analysis of bacterial lysates containing selectively 15N-labelled heterologous proteins. Our analysis permitted us to...

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Bibliographic Details
Published in:FEBS letters 2003-09, Vol.552 (2), p.207-213
Main Authors: Galvão-Botton, Leonor M.P, Katsuyama, Ângela M, Guzzo, Cristiane R, Almeida, Fábio C.L, Farah, Chuck S, Valente, Ana Paula
Format: Article
Language:English
Subjects:
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Cloning, Molecular
Gene Expression
Genes, Bacterial
HSQC, heteronuclear single quantum coherence
Molecular Structure
NMR
NMR, nuclear magnetic resonance
Nuclear Magnetic Resonance, Biomolecular - methods
Open Reading Frames
ORF, open reading frame
Protein screening
Proteomics - methods
Recombinant Proteins - chemistry
Recombinant Proteins - genetics
Solubility
Structural proteomics
Target ORF
X-ray
Xanthomonas - chemistry
Xanthomonas - genetics
Xanthomonas citri
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Summary:We applied a high-throughput strategy for the screening of targets for structural proteomics of Xanthomonas axonopodis pv citri. This strategy is based on the rapid 1H– 15N HSQC NMR analysis of bacterial lysates containing selectively 15N-labelled heterologous proteins. Our analysis permitted us to classify the 19 soluble candidates in terms of ‘foldedness’, that is, the extent to which they present a well-folded solution structure, as reflected by the quality of their NMR spectra. This classification allowed us to define a priority list to be used as a guide to select protein candidates for further structural studies.
ISSN:0014-5793
1873-3468
DOI:10.1016/S0014-5793(03)00926-8