Volume-regulated Cl- channels in human pleural mesothelioma cells |
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Authors: | Meyer Giuliano Rodighiero Simona Guizzardi Fabiana Bazzini Claudia Bottà Guido Bertocchi Cristina Garavaglia Lisa Dossena Silvia Manfredi Rosangela Sironi Chiara Catania Anna Paulmichl Markus |
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Institution: | Department of Biomolecular Sciences and Biotechnologies, University of Milan, Via Celoria 26, I-20133 Milan, Italy. giuliano.meyer@unimi.it |
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Abstract: | A novel xyloglucan-specific endo-β-1,4-glucanase (XEG), xyloglucanase, with a molecular mass of 80 kDa and a pI of 4.8, was isolated from the fungus Geotrichum sp. M128. It was found to be an endoglucanase active toward xyloglucan and not active toward carboxymethylcellulose, Avicel, or barley 1,3-1,4-β-glucan. Analysis of the precise substrate specificity using various xyloglucan oligosaccharide structures revealed that XEG has at least four subsites (−2 to +2) and specifically recognizes xylose branching at the +1 and +2 sites. The full-length cDNA encoding XEG was cloned and sequenced. It consists of a 2436-bp open reading frame encoding a 776-amino acid protein. From its deduced amino acid sequence, XEG can be classified as a family 74 glycosyl hydrolase. The cDNA encoding XEG was then expressed in Escherichia coli, and enzymatically active recombinant XEG was obtained. |
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Keywords: | Author Keywords: Xyloglucanase Xyloglucan Endo-β-1 4-glucanase |
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