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• W2023803095 abstract "The cuticle of the freshwater crayfish, Procambarus clarkii (Girard, 1852), has several extracellular, cuticular isozymes for carbonic anhydrase (CA) which were isolated and purified via affinity chromatography. To denature proteins not entombed in cuticle calcite the ground cuticle was washed twice with both a 0.5% SDS and 1.5% sodium hypochlorite solution prior to CA isolation. Total water soluble protein (WSP) was 133 ± 68 (SD) mg/100 g (6) cuticle, whereas affinity column bound protein, CA, was 191 ± 108 (SD) μg/100 g (6) cuticle. CA was 0.144% WSP. EDTA dissolved the calcium carbonate and freed the CA along with the other WSP. The CA fraction was positive to bovine CA II antibodies (ELIZA and Western Blot) and had low CA enzyme activity. The estimated molecular weights using PAGE showed one isozyme with a molecular weight of 29 kDa, which is similar to that in vertebrates, and two larger isozymes (53 and 66 kDa). All four isozymes were detected in only one sample, whereas the two larger isozymes were detected in all six cleaned cuticle samples. The larger isozymes may be dimers of the smaller isozyme and are more like those detected in oyster pearls and shell with weights of 60 kDa. La cuticule de l'ecrevisse Procambarus clarkii (Girard, 1852) contient plusieurs isoenzymes extracellulaires de l'anhydrase carbonique (CA), qui ont ete isolees et purifiees par chromatographie. Afin de denaturer les proteines non incluses dans le calcite cuticulaire, la cuticule a ete lavee deux fois a l'aide d'une solution a 0,5% de SDS et 1,5% d'hypochlorite de sodium avant l'extraction de la CA. Les proteines totales solubles dans l'eau (WSP) representaient 133 ± 68 (SD) mg/100 g (6) de cuticule, tandis que la proteine associee CA, etait de 191 ± 108 (SD) μg/100 g (6) de cuticule, soit CA = 0,144% WSP. L'EDTA a dissous le carbonate de calcium et libere la CA avec les autres WSP. La fraction de CA s'est revelee positive aux anticorps bovins CA II (ELISA et Western Blot) et a une activite enzymatique CA detectable. Les poids moleculaires estimees en utilisant PAGE, ont revele une isoenzyme avec un poids moleculaire de 29 kDa, qui est analogue a celle trouvee chez les Vertebres, et deux plus grands isoenzymes (53 et 66 kDa). Les quatre isoenzymes ont ete detectees seulement dans un echantillon tandis que les deux plus grandes ont ete detectees dans tous les six echantillons de cuticule laves. Les plus grandes pourraient etre des dimeres de la plus petite et sont comparables a celles detectees dans les perles et la coquille d'huitre avec des poids moleculaires de 60 kDa." @default.
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• W2023803095 date "2002-01-01" @default.
• W2023803095 modified "2023-09-27" @default.
• W2023803095 title "CARBONIC ANHYDRASE IN MINERALIZATION OF THE CRAYFISH CUTICLE" @default.
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• W2023803095 doi "https://doi.org/10.1163/156854002763270473" @default.
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