Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin

Hemocyanin of the spider <i>Polybetes pythagoricus</i>, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters K<sub>m</sub> and V<sub&...

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Autores principales: Laino, Aldana, Lavarías, Sabrina María Luisa, Suárez, Gustavo, Lino, Agustina Berenice, Cunningham, Mónica Liliana
Formato: Articulo Preprint
Lenguaje:Inglés
Publicado: 2015
Materias:
Ciencias Exactas
Ciencias Naturales
Polybetes pythagoricus
Hemocyanin
Phenoloxidase Activity
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/127717
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-127717
record_format dspace
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Exactas
Ciencias Naturales
Polybetes pythagoricus
Hemocyanin
Phenoloxidase Activity
spellingShingle Ciencias Exactas
Ciencias Naturales
Polybetes pythagoricus
Hemocyanin
Phenoloxidase Activity
Laino, Aldana
Lavarías, Sabrina María Luisa
Suárez, Gustavo
Lino, Agustina Berenice
Cunningham, Mónica Liliana
Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin
topic_facet Ciencias Exactas
Ciencias Naturales
Polybetes pythagoricus
Hemocyanin
Phenoloxidase Activity
description Hemocyanin of the spider <i>Polybetes pythagoricus</i>, in addition to its typical role as an oxygen transporter, also exhibits a phenoloxidase activity induced by micellar concentrations of SDS. In the present work, we found the kinetic parameters K<sub>m</sub> and V<sub>max</sub> of <i>Polybetes pythagoricus</i> hemocyanin (<i>Pp</i>Hc) PO activity to be 0.407mM and 0.081 μmolmin⁻¹mg protein⁻¹, respectively. Dopamine was used as the substrate with SDS at a final concentration of 10mM and a 30-min incubation at 25°C. Conformational changes in Hc associated with the SDS treatment were analyzed using far-UV circular dichroism, intrinsic fluorescence and absorption spectroscopy. The secondary and tertiary structural changes of <i>Pp</i>Hc induced by SDS led to increases in α-helical content and tryptophan fluorescence intensity. A reduction in the absorption spectrum at 340 nm in the presence of SDS was also observed. These results suggest that the SDS-induced PO activity of <i>Pp</i>Hc can be ascribed to conformational changes in the local environment of the typer-3 copper active site.
format Articulo
Preprint
author Laino, Aldana
Lavarías, Sabrina María Luisa
Suárez, Gustavo
Lino, Agustina Berenice
Cunningham, Mónica Liliana
author_facet Laino, Aldana
Lavarías, Sabrina María Luisa
Suárez, Gustavo
Lino, Agustina Berenice
Cunningham, Mónica Liliana
author_sort Laino, Aldana
title Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin
title_short Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin
title_full Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin
title_fullStr Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin
title_full_unstemmed Characterization of Phenoloxidase Activity From Spider <i>Polybetes pythagoricus</i> Hemocyanin
title_sort characterization of phenoloxidase activity from spider <i>polybetes pythagoricus</i> hemocyanin
publishDate 2015
url http://sedici.unlp.edu.ar/handle/10915/127717
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