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 (1)H-(15)N HSQC NMR analysis of bacterial lysates containing selectively (15)N-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.
Galvão-Botton
Leonor M P
LM
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, CEP 05508-900, SP, São Paulo, Brazil.
Katsuyama
Angela M
AM
Guzzo
Cristiane R
CR
Almeida
Fábio C L
FC
Farah
Chuck S
CS
Valente
Ana Paula
AP
D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics*; D003001:Cloning, Molecular; D015870:Gene Expression; D005798:Genes, Bacterial; D015394:Molecular Structure; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D016366:Open Reading Frames; D040901:Proteomics / Q000379:methods*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012995:Solubility; D014974:Xanthomonas / Q000737:chemistry* / Q000235:genetics*
D001426:Bacterial Proteins; D011994:Recombinant Proteins