Glycoforms of beta-lactoglobulin with improved thermostability and preserved structural packing.
In this article we show how various degrees of glycosylation can be used to control the thermal stability of proteins. The primary amines of beta-lactoglobulin were glycosylated with glucose or fructose within a range of non-denaturing reaction parameters. The modified fractions were characterized and analyzed for structural stability and hydrophobic exposure. The modification procedure gave rise to the production of glycoproteins with a well-defined Gaussian distribution, where glucose appeared more reactive than fructose. The integrity of the secondary, tertiary, and quaternary structures remained unaffected by the modification procedure. However, upon heating the stability of the modified fractions increased up to 6 K. Here we demonstrate the effects on the thermodynamic properties of proteins by glycosylation; this work serves as a first step in understanding and controlling the process underlying aggregation of glycosylated proteins.
Wageningen Centre for Food Sciences, Diedenweg 20, P.O. Box 557, 6700 AN Wageningen, The Netherlands.
Kerensa
K
Broersen
Alphons G J
AG
Voragen
Rob J
RJ
Hamer
Harmen H J
HH
De Jongh
D005632:Fructose / Q000737:chemistry*; D005947:Glucose / Q000737:chemistry*; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D007782:Lactoglobulins / Q000737:chemistry*; D046911:Macromolecular Substances; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D020033:Protein Isoforms / Q000737:chemistry; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011961:Receptors, Fc; D013696:Temperature
C045281:IgA receptor; D007782:Lactoglobulins; D046911:Macromolecular Substances; D020033:Protein Isoforms; D011961:Receptors, Fc; D005632:Fructose; D005947:Glucose