Glycoprotein-surfactant interactions: A calorimetric and spectroscopic investigation of the phytase-SDS system

被引:28
作者
Bagger, Heidi L.
Hoffmann, Soren V.
Fuglsang, Claus C.
Westh, Peter
机构
[1] Roskilde Univ, Dept Sci Syst & Models, DK-4000 Roskilde, Denmark
[2] Aarhus Univ, Inst Storage Ring Facil, DK-8000 Aarhus, Denmark
[3] Novozymes AS, DK-2880 Bagsvaerd, Denmark
关键词
surfactant-protein interactions; thermodynamics; glycosylation; stability; circular dichroism; titration calorimetry; SODIUM DODECYL-SULFATE; BOVINE SERUM-ALBUMIN; ISOTHERMAL TITRATION CALORIMETRY; HUMICOLA-INSOLENS CUTINASE; PENIOPHORA-LYCII PHYTASE; ANIONIC SURFACTANT; CIRCULAR-DICHROISM; BINDING ISOTHERM; HORSERADISH-PEROXIDASE; KINETIC STABILITY;
D O I
10.1016/j.bpc.2007.06.005
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 interactions of sodium dodecyl sulfate (SDS) and two glyco-variants of the enzyme phytase from Peniophora lycii were investigated. One variant (Phy) was heavily glycosylated while the other (dgPhy) was enzymatically deglycosylated. Effects at 24 degrees C of titrating SDS to Phy and dgPhy were studied by Isothermal Titration Calorimetry (ITC) and Synchrotron Radiation Circular Dichroism (SRCD) spectroscopy. Comparisons of results for the two variants were used to elucidate glycan-surfactant interrelationships. The CD spectra suggested that both the native and the SDS-denatured states of the two variants were mutually similar, and hence that the denaturation process was structurally equivalent for the two glyco-variants. The denatured state was far from fully unfolded and probably retained a substantial content of native-like structure. Furthermore, it was found that the glycans brought about only a small increase in the resistance towards SDS induced denaturation. The SDS concentration required to denature half of the protein molecules differed less than 1 MM for the two variants. The affinity for SDS of both variants was unusually low. The amount of bound SDS (w/w) at different stages of the binding isothenn was 310 times lower than that reported for the most previously investigated globular proteins. Analysis of the relative affinity of the glycan and peptide moieties suggested that the carbohydrates bind much less surfactant. At saturation, glycans adsorbed about half as much SDS (in g/g) as the peptide moiety of Phy and about five times less than average proteins. (c) 2007 Elsevier B.V. All rights reserved.
引用
收藏
页码:251 / 258
页数:8
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