Identification of the active site residues in ATP-citrate lyase's carboxy-terminal portion
| Autor: | Vinh H Nguyen, Ana Medina, Marie E. Fraser, Isabel Usón, Noreen Singh |
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| Přispěvatelé: | Natural Sciences and Engineering Research Council of Canada, Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), Ministerio de Ciencia, Innovación y Universidades (España), Argonne National Laboratory (US), Canadian Institutes of Health Research, National Research Council of Canada, University of Saskatchewan, Canada Foundation for Innovation |
| Rok vydání: | 2019 |
| Předmět: |
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Molecular ATP citrate lyase CoA‐binding Full‐Length Papers education Reductive tricarboxylic acid cycle Chlorobium Crystallography X-Ray Biochemistry 03 medical and health sciences Citrate synthase Molecular Biology 030304 developmental biology chemistry.chemical_classification 0303 health sciences Chymotrypsin Binding Sites biology Chemistry 030302 biochemistry & molecular biology Active site Lyase Site‐directed mutagenesis Citric acid cycle Enzyme Proteolysis biology.protein ATP Citrate (pro-S)-Lyase Biocatalysis Mutagenesis Site-Directed |
| Zdroj: | Protein Sci Digital.CSIC. Repositorio Institucional del CSIC instname |
| ISSN: | 1469-896X |
| Popis: | ATP‐citrate lyase (ACLY) catalyzes production of acetyl‐CoA and oxaloacetate from CoA and citrate using ATP. In humans, this cytoplasmic enzyme connects energy metabolism from carbohydrates to the production of lipids. In certain bacteria, ACLY is used to fix carbon in the reductive tricarboxylic acid cycle. The carboxy(C)‐terminal portion of ACLY shows sequence similarity to citrate synthase of the tricarboxylic acid cycle. To investigate the roles of residues of ACLY equivalent to active site residues of citrate synthase, these residues in ACLY from Chlorobium limicola were mutated, and the proteins were investigated using kinetics assays and biophysical techniques. To obtain the crystal structure of the C‐terminal portion of ACLY, full‐length C. limicola ACLY was cleaved, first non‐specifically with chymotrypsin and subsequently with Tobacco Etch Virus protease. Crystals of the C‐terminal portion diffracted to high resolution, providing structures that show the positions of active site residues and how ACLY tetramerizes. Natural Sciences and Engineering Research Council of Canada, Grant/Award Number: Discovery Grant; Spanish Ministry of Science and Innovations, Grant/Award Numbers: BES‐2017‐080368, BIO2015‐64216‐P, MDM2014‐0435; Argonne National Laboratory, Grant/Award Number: DE‐AC02‐06CH11357; Canadian Institutes of Health Research; National Research Council Canada; Western Economic Diversification Canada; Government of Saskatchewan; University of Saskatchewan; Natural Sciences and Engineering Research Council of Canada (NSERC); Canada Foundation for Innovation; Queen Elizabeth II Scholarship; Canada Graduate Scholarship—Master's Program; NSERC |
| Databáze: | OpenAIRE |
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