Data/hora: 24/06/2024 19:48:35

Biblioteca(s): | Embrapa Agricultura Digital.
Data corrente: | 05/10/2011
Data da última atualização: | 06/01/2012
Autoria: | GHELFI, A.; GAZIOLA, S. A.; CIA, M. C.; CHABREGAS, S. M.; FALCO, M. C.; KUSER-FALCÃO, P. R.; AZEVEDO, R. A.
Afiliação: | A. GHELFI, Universidade Federal do Amazonas; S. A. GAZIOLA, ESALQ/USP; MARIANA CICARELLA CIA, ESALQ/USP; SABRINA M. CHABREGAS, Centro de Tecnologia Canavieira; M. C. FALCO, Centro de Tecnologia Canavieira; PAULA REGINA KUSER-FALCÃO, CNPTIA; R. A. AZEVEDO, ESALQ/USP.
Título: | Cloning, expression, molecular modelling and docking analysis of glutathione transferase from Saccharum officinarum.
Ano de publicação: | 2011
Fonte/Imprenta: | Annals of Applied Biology, Cambridge, v. 159, n. 2, p. 267-280, 2011.
DOI: | 10.1111/j.1744-7348.2011.00491.x
Idioma: | Inglês
Conteúdo: | Sugarcane yield and quality are affected by a number of biotic and abiotic stresses. In response to such stresses, plants may increase the activities of some enzymes such as glutathione transferase (GST), which are involved in the detoxification of xenobiotics. Thus, a sugarcane GST was modelled and molecular docked using the program LIGIN to investigate the contributions of the active site residues towards the binding of reduced glutathione (GSH) and 1-chloro-2,4-dinitrobenzene (CDNB). As a result, W13 and I119 were identified as key residues for the specificity of sugarcane GSTF1 (SoGSTF1) towards CDNB. To obtain a better understanding of the catalytic specificity of sugarcane GST (SoGSTF1), two mutants were designed, W13L and I119F. Tertiary structure models and the same docking procedure were performed to explain the interactions between sugarcane GSTs with GSH and CDNB. An electron-sharing network for GSH interaction was also proposed. The SoGSTF1 and the mutated gene constructions were cloned and expressed in Escherichia coli and the expressed protein purified. Kinetic analyses revealed different Km values not only for CDNB, but also for GSH. The Km values were 0.2, 1.3 and 0.3 mM for GSH, and 0.9, 1.2 and 0.5 mM for CDNB, for the wild type, W13L mutant and I119F mutant, respectively. The Vmax values were 297.6, 224.5 and 171.8 umol min-1 mg-1 protein for GSH, and 372.3, 170.6 and 160.4 umol min-1 mg-1 protein for CDNB.
Palavras-Chave: | Cana-de-açúcar; Clonagem molecular; Modelagem molecular.
Thesagro: | Saccharum Officinarum.
Thesaurus Nal: | Glutathione transferase; Molecular cloning; Molecular models; Sugarcane.
Categoria do assunto: | G Melhoramento Genético
Registro original: | Embrapa Agricultura Digital (CNPTIA)
Exemplares: | CNPTIA - (15977-1) - UPC - AP - DD -  -  -  - DIS