Refolding and reactivation of calf intestinal alkaline phosphatase with excess magnesium ions.
Int J Biochem Cell Biol
; 34(10): 1241-7, 2002 Oct.
Article
in En
| MEDLINE
| ID: mdl-12127574
ABSTRACT
It is well known that Mg(2+) is an essential component in many biological processes. This research investigated the courses of both the reactivation and the refolding in the absence and presence of Mg(2+) ions. Calf intestinal alkaline phosphatase (CIP) was extensively denatured in 3 M guanidine hydrochloride (GdnHCl) solution for 2 h. Under suitable renaturation conditions, about 60-70% of the activity was recovered in the absence and presence of different magnesium ion concentrations. The refolding processes followed two-phase courses, whereas the reactivation processes were monophasic after dilution in proper solutions with or without Mg(2+). The magnesium ions affected both the reactivation and the refolding courses of unfolded CIP. A comparison of rate constants for the refolding of unfolded CIP with those for recovery of enzyme activity at different Mg(2+) concentrations showed that they were not synchronized. The activity recovery was speeded up due to the presence of Mg(2+) ions; while the refolding course of unfolded CIP was somewhat inhibited by the excess Mg(2+).
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Protein Folding
/
Alkaline Phosphatase
/
Magnesium
/
Antigens, Neoplasm
Limits:
Animals
Language:
En
Journal:
Int J Biochem Cell Biol
Journal subject:
BIOQUIMICA
Year:
2002
Type:
Article