DOI | Trouver le DOI : https://doi.org/10.1021/bi00224a008 |
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Auteur | Rechercher : Labruyère, Elisabeth; Rechercher : Mock, Michèle; Rechercher : Surewicz, Witold K.1; Rechercher : Mantsch, Henry H.1; Rechercher : Rose, Thierry; Rechercher : Munier, Hélène; Rechercher : Sarfati, Robert S.; Rechercher : Bârzu, Octavian |
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Affiliation | - Conseil national de recherches du Canada. Institut Steacie des sciences moléculaires du CNRC
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Format | Texte, Article |
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Résumé | A truncated, 541-residue-long, Bacillus anthracis adenylate cyclase was expressed in Escherichia coli. The purified protein (CYA 62) exhibited catalytic and CaM-binding properties identical with those of the wild-type enzyme secreted by B. anthracis. The analysis of the secondary structure of the CYA 62 protein by Fourier transform infrared spectroscopy and circular dichroism revealed the dominance of β-type structure. The protein shows a relatively low thermal stability with the midpoint denaturation temperature at 45ºC. A catalytically inactive varient of CYA 62 in which Gln substituted for Lys-346 (CYA 62 K346Q) was comparatively analyzed for its secondary structure and thermal stability, as well as ligand-binding properties with fluorescent derivaties of ATP and calmodulin. The K346Q variant of CYA 62 has a similar secondary structure and comparable calmodulin binding porperties to those of the parent protein and exhibits only slightly reduced thermal stability (the apparent midpoint denaturation temperature is at 43ºC). Despite these similarities, the binding of 3'-anthraniloyl-2'-deoxy-ATP (a fluorescent ATP analogue) to the modified protein is severely impaired, from which we conclude that the prime function of Lys-346 in the wild-type enzyme from B. anthracis is to ensure tight binding of the nucleotide substrate to the active site. |
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Date de publication | 1991-03 |
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Dans | |
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Langue | anglais |
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Publications évaluées par des pairs | Oui |
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Numéro du CNRC | NRCC 32284 |
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Numéro NPARC | 23001494 |
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Exporter la notice | Exporter en format RIS |
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Signaler une correction | Signaler une correction (s'ouvre dans un nouvel onglet) |
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Identificateur de l’enregistrement | c1e88bb2-cebd-4347-a225-2852d6d56c1e |
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Enregistrement créé | 2017-02-20 |
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Enregistrement modifié | 2020-03-17 |
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