Expression, purification, enzymatic characterization and crystallization of glyceraldehyde-3-phosphate dehydrogenase from Naegleria gruberi, the first one from phylum Percolozoa
| Autor: | Agnes Thiane Pereira Machado, Jorge Iulek, Marcio Silva |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Protozoan Proteins Gene Expression Crystallography X-Ray Genome 03 medical and health sciences Protein Domains parasitic diseases Escherichia coli Enzyme kinetics Glyceraldehyde 3-phosphate dehydrogenase chemistry.chemical_classification Naegleria fowleri 030102 biochemistry & molecular biology biology Naegleria gruberi Glyceraldehyde-3-Phosphate Dehydrogenases Naegleria biology.organism_classification Recombinant Proteins Metabolic pathway 030104 developmental biology Enzyme Biochemistry chemistry biology.protein NAD+ kinase Biotechnology |
| Zdroj: | Protein expression and purification. 127 |
| ISSN: | 1096-0279 |
| Popis: | Naegleria gruberi had its genome sequenced by Fritz-Laylin and collaborators in 2010. It is not pathogenic, but has characteristics similar to those of Naegleria fowleri, opportunistic pathogen that can cause fatal encephalitis in humans. N. gruberi genome has contributed to a better understanding of the primitive eukaryotic metabolism and revealed the complexity of several metabolic pathways. In this paper we describe the expression, purification, enzyme characterization and crystallization of N. gruberi GAPDH, the first one for an organism belonging to phylum Percolozoa. The results indicated that 10 mM, 8.0 and 25 °C are the optimum arsenate concentration, pH and temperature, respectively. The enzyme presents allosteric positive cooperativity for substrates NAD+ and G3P as indicated by the Hill coefficients. The phylogenetic proximity between N. fowleri and N. gruberi suggests that contributions from the study of the latter might provide information to assist the search for treatments of Primary Amebic Meningoencephalitis, especially, in this work, taking into account that GAPDH is identified as a therapeutic target. |
| Databáze: | OpenAIRE |
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