Susceptibility towards intramolecular disulphide-bond formation affects conformational stability and folding of human basic fibroblast growth factor

被引：33

作者：

Estapé, D ^{[1
]}

van den Heuvel, J ^{[1
]}

Rinas, U ^{[1
]}

机构：

[1] GBF Natl Res Ctr Biotechnol, Div Biochem Engn, D-38124 Braunschweig, Germany

来源：

BIOCHEMICAL JOURNAL | 1998年 / 335卷

关键词：

D O I：

10.1042/bj3350343

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：

The conformational stability and the folding properties of the all-beta-type protein human basic fibroblast growth factor (hFGF-2) were studied by means of fluorescence spectroscopy. The results show that the instability of the biological activity of hFGF-2 is also reflected in a low conformational stability of the molecule. The reversibility of the unfolding and refolding process was established under reducing conditions. Determination of the free-energy of unfolding in the presence of reducing agents revealed that the conformational stability of hFGF-2 (Delta G(app)(H2O) congruent to 21 kJ.mol-(1,) 25 degrees C) is low compared with other globular proteins under physiological conditions (20-60 kJ.mol(-1)). However, the conformational stability of hFGF-2 is particularly low under non-reducing conditions. This instability is attributed to intramolecular disulphide-bond formation, rendering the molecule more susceptible to denaturant-induced unfolding. In addition, denaturant-induced unfolding of hFGF-2 renders the protein more susceptible to irreversible oxidative denaturation. Experimental evidence is provided that the irreversibility of the unfolding and refolding process in the absence of reducing agents is linked to the formation of an intramolecular disulphide bond involving cysteines 96 and 101.

引用

页码：343 / 349

页数：7

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