High‐throughput screening of structural proteomics targets using NMR

FEBS Letters

Galvão-Botton, Leonor M.P²  Guzzo, Cristiane R²  Katsuyama, Ângela M²  Valente, Ana Paula¹  Almeida, Fábio C.L¹  Farah, Chuck S² 

[1] Departamento de Bioquı́mica Médica, ICB, CCS, Universidade Federal do Rio de Janeiro, CEP 21941-590, Rio de Janeiro, RJ, Brazil;Departamento de Bioquı́mica, Instituto de Quı́mica, Universidade de São Paulo, CEP 05508-900, São Paulo, SP, Brazil

关键词: Structural proteomics;    NMR;    X-ray;    Protein screening;    Target ORF;    Xanthomonas citri;    ORF;    open reading frame;    NMR;    nuclear magnetic resonance;    HSQC;    heteronuclear single quantum coherence;   

DOI  :  10.1016/S0014-5793(03)00926-8

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

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.

【 授权许可】

Unknown