On the temperature stability of extracellular hemoglobin of Glossoscolex paulistus, at different oxidation states: SAXS and DLS studies

被引:16
作者
Carvalho, Jose Wilson P. [1 ]
Santiago, Patricia S. [1 ,2 ]
Batista, Tatiana [1 ]
Garrido Salmon, Carlos Ernesto [3 ]
Barbosa, Leandro R. S. [4 ]
Itri, Rosangela [4 ]
Tabak, Marcel [1 ]
机构
[1] Univ Sao Paulo, Inst Quim Sao Carlos, Sao Carlos, SP, Brazil
[2] Univ Estadual Paulista, Registro, SP, Brazil
[3] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Fis, Ribeirao Preto, SP, Brazil
[4] Univ Sao Paulo, Inst Fis, BR-01498 Sao Paulo, Brazil
基金
巴西圣保罗研究基金会;
关键词
Extracellular hemoglobin; Glossoscolex paulistus; Oligomeric dissociation; Thermal stability; DLS; SAXS; LUMBRICUS-TERRESTRIS HEMOGLOBIN; X-RAY-SCATTERING; STRUCTURAL-ANALYSIS; PH; RESOLUTION; SUBUNITS;
D O I
10.1016/j.bpc.2012.02.004
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 degrees C, and R-g is 108 angstrom, independent of the oxidation form. At pH 7.0, protein denaturation and aggregation occurs above 55 degrees C and 60 degrees C, for oxy and met-HbGp, respectively. Cyanomet-HbGp, at pH 7.0, is stable up to 60 degrees C. At alkaline pH (8.0-9.0) and higher temperature, an irreversible dissociation process is observed, with a decrease of R-g, D-max 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. (C) 2012 Elsevier B.V. All rights reserved.
引用
收藏
页码:44 / 55
页数:12
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