Glycosylation and thermodynamic versus kinetic stability of horseradish peroxidase

FEBS Letters

Tams, Jeppe W¹  Welinder, Karen G¹ 

[1] Department of Protein Chemistry, Institute of Molecular Biology, University of Copenhagen, Øster Farimagsgade 2A, DK-1353 Copenhagen K, Denmark

关键词: Glycoprotein stability;    Glycoprotein unfolding;    Horseradish peroxidase;    Thermodynamic stability;    Kinetic stability;    GdmCl;    guanidinium chloride;    G*;    standard molar transition state free energy;    G°;    standard molar free energy;    HRP;    horseradish peroxidase isoenzyme C;    d-HRP;    deglycosylated homogeneous HRP;   

DOI  :  10.1016/S0014-5793(97)01573-1

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The influence of N-linked glycans on the stability of glycoproteins has been studied using horseradish peroxidase isoenzyme C (HRP), which contains eight asparagine-linked glycans. HRP was deglycosylated (d-HRP) with trifluoromethanesulfonic acid and purified to an enzymatically active homogeneous protein containing (GlcNAc)₂ glycans. The thermal stability of HRP and d-HRP at pH 6.0, measured by residual activity, was indistinguishable and showed transition midpoints at 57°C, whereas the unfolding in guanidinium chloride at pH 7.0, 23°C was 2–3-fold faster for d-HRP than for HRP. The results are compatible with a glycan-induced decrease in the dynamic fluctuation of the polypeptide chain.

【 授权许可】

Unknown