Stabilization of Ribonuclease B Activity by Concentrated Xylose Solutions

Ribonuclease B has become a paradigm as a simple example of an N-linked glycoprotein. We have found that certain affinity-purified preparations of this enzyme demonstrated a pronounced tendency to lose activity if stored as dilute aqueous solutions. Such inactivation is accelerated by the presence o...

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Bibliographic Details
Published in:Biochemical and biophysical research communications 1995-02, Vol.207 (1), p.432-437
Main Authors: Williams, G.A., Macevilly, U., Ryan, R., Harrington, M.G.
Format: Article
Language:English
Subjects:
Chromatography, Affinity
Enzyme Stability
Kinetics
Methylmannosides - pharmacology
Ribonucleases - chemistry
Ribonucleases - isolation & purification
Ribonucleases - metabolism
Solutions
Spectrophotometry, Ultraviolet
Xylose - pharmacology
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Summary:Ribonuclease B has become a paradigm as a simple example of an N-linked glycoprotein. We have found that certain affinity-purified preparations of this enzyme demonstrated a pronounced tendency to lose activity if stored as dilute aqueous solutions. Such inactivation is accelerated by the presence of NaCl, but can be counteracted by inclusion of high (1 mol/l) concentrations of xylose. Enzyme activity cannot be restored by addition of xylose after storage of the enzyme. In marked contrast to α-methyl-mannoside, xylose does not prevent ribonuclease B from binding to concanavalin A and so may be used to stabilize the enzyme during purification by lectin affinity chromatography.
ISSN:0006-291X
1090-2104
DOI:10.1006/bbrc.1995.1206