期刊
PROTEIN EXPRESSION AND PURIFICATION
卷 182, 期 -, 页码 -出版社
ACADEMIC PRESS INC ELSEVIER SCIENCE
DOI: 10.1016/j.pep.2021.105838
关键词
Metallothionein zinc-binding proteins; Purification; Secondary structure; Anti-oxidant activity
类别
资金
- China's national key RD plan [1110XJ8019477]
Zinc-binding proteins named MT-M-I and MT-M-II were purified from metal-exposed hairy clams and found to have similar molecular weights, capable of binding 6 and 7 equivalents of Zn2+ in vitro. Characterized by their migration behaviors in Tricine-SDS-PAGE and amino acid compositions, these proteins conform to the molecular characteristics of metallothionein proteins. Additionally, MT-M-I exhibited stronger antioxidant activities compared to MT-M-II.
Zinc-binding proteins named MT-M-I and MT-M-II were obtained after purification from metal-exposed hairy clams (Arca subcrenata) using gel permeation and ion-exchange chromatography. MT-M-I and MT-M-II were resolved by ion-exchange chromatography, and they were found to have similar molecular weights. MT-M-I and MT-M-II can bind 6 and 7 equivalents of Zn2+ in vitro, and they showed unusual migration behaviors in Tricine sodium dodecyl sulfate polyacrylamide gel electrophoresis (Tricine-SDS-PAGE). Such migration behaviors may be due to themetal thiolate clusters in these proteins. In terms of amino acid composition, the proportion of cysteine in MT-M-I and MT-M-II was approximately 30%, and glycine accounted for approximately 15%, where as aromatic amino acids were absent. Considering the performance in Tricine-SDS-PAGE and the amino acid compositions, MT-M-I and MT-M-II conform to the molecular characteristics of the metallothionein proteins. The structures were explored using circular dichroism (CD) and Fourier-transform infrared spectroscopy (FTIR). Also determined the antioxidant activities in terms of DPPH radical scavenging ability, hydroxyl radical ((OH)-O-center dot) scavenging ability, and ferric-reducing/antioxidant power. The antioxidant activities of MT-M-I were found to be stronger than those of MT-M-II.
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