Expression, purification, and circular dichroism analysis of human CDK9

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Leopoldino, Andreia Machado ; Canduri, Fernanda ; Cabral, Hamilton ; Junqueira, Magno ; Marqui, Alessandra Bernadete Trovó de ; Apponi, Luciano H. ; Fonseca, Isabel Osório da ; Domont, Gilberto Barbosa ; Santos, Diógenes S. ; Valentini, Sandro ; et al.

• Tipo de documento: Artigo Científico
• Fonte: Protein Expression and Purification; v. 47, n. 2, p. 614-620, June 2006.
• Área do conhecimento: Ciências Biológicas - Genética
• Assunto(s): Neoplasias AIDS

Resumo

The human cyclin-dependent kinase 9 (CDK9) protein was expressed in E. coli BL21 using the pET23a vector at 30 °C. Several milligrams of protein were purified from soluble fraction using ionic exchange and ATP-affinity chromatography. The structural quality of recombinant CDK9 and the estimation of its secondary structure were obtained by circular dichroism. Structural models of CDK9 presented 26% of helices in agreement with the spectra by circular dichroism analysis. This is the first report on human CDK9 expression in Escherichia coli and structure analysis and provides the first step for the development of CDK9 inhibitors. (AU)

• Processo FAPESP: 01/07532-0 - Structural genomics of cyclin dependent kinases and plant defensive proteinases and their natural inhibitors
• Beneficiário: Walter Filgueira de Azevedo Junior
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 02/09388-7 - Estudo de biomarcadores proteicos em câncer de cabeça e pescoço
• Beneficiário: Andréia Machado Leopoldino
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 02/04383-7 - Estudo estrutural das enzimas da via metabólica do ácido chiquímico
• Beneficiário: Walter Filgueira de Azevedo Junior
• Modalidade de apoio: Auxílio à Pesquisa - Regular