Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8326885 | International Journal of Biological Macromolecules | 2018 | 8 Pages |
Abstract
To gain insight into the effect of Cu2+ on the activity and structure of alkaline phosphatase (ALP) from Macrobrachium rosenbergii, the enzyme was purified using ammonium sulfate fractionation, Sephacryl S-200, and DEAE anion exchange chromatography. We studied Cu2+-mediated inhibition and aggregation of ALP, and found that Cu2+ significantly inactivated ALP activity with an IC50 of 1.47â¯Â±â¯0.02â¯mM. We further revealed that Cu2+ reversibly inhibited ALP in a mixed-type manner with Kiâ¯=â¯0.41â¯Â±â¯0.02â¯mM. Time-interval kinetics showed that the inhibition followed first-order reaction kinetics. This process was associated with conformational changes and significant transient free-energy change. Spectrofluorometry results showed that Cu2+ induced ALP tertiary structural changes, including the exposure of hydrophobic surfaces that directly induced ALP aggregation. The results provide new information regarding ALP from M. rosenbergii.
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Authors
Zhi-Jiang Wang, Wenming Ma, Jun-Mo Yang, Yani Kang, Yong-Doo Park,