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Biophys Chem ; 163-164: 44-55, 2012 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-22397813

RESUMEN

Glossoscolex paulistus hemoglobin (HbGp) was studied by dynamic light scattering (DLS) and small angle X-ray scattering (SAXS). DLS melting curves were measured for met-HbGp at different concentrations. SAXS temperature studies were performed for oxy-, cyanomet- and met-HbGp forms, at several pH values. At pH 5.0 and 6.0, the scattering curves are identical from 20 to 60 °C, and Rg is 108 Å, independent of the oxidation form. At pH 7.0, protein denaturation and aggregation occurs above 55 °C and 60 °C, for oxy and met-HbGp, respectively. Cyanomet-HbGp, at pH 7.0, is stable up to 60 °C. At alkaline pH (8.0-9.0) and higher temperature, an irreversible dissociation process is observed, with a decrease of Rg, Dmax and I(0). Analysis by p(r), obtained from GNOM, and OLIGOMER, was used to fit the SAXS experimental scattering curves by a combination of theoretical curves obtained for HbLt fragments from the crystal structure. Our results show clearly the increasing contribution of smaller molecular weight fragments, as a function of increasing pH and temperature, as well as, the order of thermal stabilities: cyanomet->oxy->met-HbGp.


Asunto(s)
Hemoglobinas/química , Luz , Oligoquetos/metabolismo , Dispersión de Radiación , Dispersión del Ángulo Pequeño , Difracción de Rayos X , Animales , Hemoglobinas/metabolismo , Concentración de Iones de Hidrógeno , Hierro/química , Metahemoglobina/análogos & derivados , Metahemoglobina/química , Metahemoglobina/metabolismo , Oxidación-Reducción , Estabilidad Proteica , Temperatura
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