Self-association of human beta-galactocerebrosidase: Dependence on pH, salt, and surfactant


Autoři: Eunhee Lee aff001;  Nazila Salamat-Miller aff001;  Walter F. Stafford aff002;  Katherine Taylor aff001
Působiště autorů: Process Development and Technical Services, Shire US Inc., a Takeda company, Lexington, MA, United States of America aff001;  Cardiovascular Biology Program and AUC Research Laboratory, Boston Biomedical Research Institute, Watertown, MA, United States of America aff002
Vyšlo v časopise: PLoS ONE 14(12)
Kategorie: Research Article
doi: 10.1371/journal.pone.0226618

Souhrn

Krabbe disease, also known as globoid cell leukodystrophy, is a rare genetic neurodegenerative disease caused by a deficiency of the galactocerebrosidase enzyme. To understand the association status of human beta-galactocerebrosidase (hGALC) in solution, we employed analytical ultracentrifugation (AUC). Our AUC results show that hGALC has a tendency for reversible self-association. Self-association decreases as the concentration of sodium chloride increases from 50 to 500 mM. This indicates that ionic interactions are involved in the association. The association is also dependent on pH, and high order oligomerization decreases as the pH increases from 4.5 to 7.5. Taken together, our results indicate that hGALC has the highest tendency for oligomerization at physiological ionic strength and pH (lysosomal lumen). This is the first report describing the self-associating property of hGALC in solution.

Klíčová slova:

Enzyme purification – Enzyme replacement therapy – Lysosomes – Sedimentation – Sodium chloride – Sodium phosphate – Viscosity – Recombinant protein purification


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Článek vyšel v časopise

PLOS One


2019 Číslo 12