Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i>
We report here the purification of a novel metal-binding protein from <i>Oratosquilla oratoria</i> (<i>O. oratoria</i> MT-1) by gel and ion-exchange chromatography. SDS-PAGE and MALDI-TOF analyses demonstrated that isolated <i>O. oratoria</i> MT-1 was of high puri...
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MDPI AG
2022-04-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/7/2320 |
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| author | Guang-Ming Mei Xiao-Hua Wu Xiao-Jun Zhang Jie Gu Yi Fang Chun-Ying Meng Wen-Ge Yang |
| author_facet | Guang-Ming Mei Xiao-Hua Wu Xiao-Jun Zhang Jie Gu Yi Fang Chun-Ying Meng Wen-Ge Yang |
| author_sort | Guang-Ming Mei |
| collection | DOAJ |
| description | We report here the purification of a novel metal-binding protein from <i>Oratosquilla oratoria</i> (<i>O. oratoria</i> MT-1) by gel and ion-exchange chromatography. SDS-PAGE and MALDI-TOF analyses demonstrated that isolated <i>O. oratoria</i> MT-1 was of high purity with a molecular weight of 12.4 kDa. The fluorescence response to SBD-F derivatives revealed that <i>O. oratoria</i> MT-1 contained a large number of sulfhydryl groups, which is a general property of metallothioneins. Zn and Cu metal stoichiometries for <i>O. oratoria</i> MT-1 were 3.97:1 and 0.55:1, respectively. The proportion of cysteine (Cys) residues in the amino acid composition was 32.69%, and aromatic amino acids were absent. The peptide sequence coverage with <i>Macrobrachium rosenbergii</i> calmodulin (accession AOA3S8FSK5) was 60%. Infrared spectroscopy of <i>O. oratoria</i> MT-1 revealed two obvious peaks at absorption frequencies for the amide I band and the amide II band. CD spectra revealed that the secondary structure was mainly composed of random coil (57.6%) and β-sheet (39.9%). An evaluation of in vitro antioxidant activity revealed that isolated <i>O. oratoria</i> MT-1 has strong reducing activities, exhibiting scavenging rates for DPPH and OH of 77.8% and 75.8%, respectively (IC<sub>50</sub> values 0.57 mg/mL and 1.1 mg/mL). <i>O. oratoria</i> MT-1 may be used as a functional additive in cosmetics, health foods, and medical products, as well as a reference material for quantitative analysis of metallothionein in such products. |
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| last_indexed | 2024-03-09T11:36:01Z |
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| spelling | doaj.art-896f63b8d00d4b49a2f401b90ec6516e2023-11-30T23:42:37ZengMDPI AGMolecules1420-30492022-04-01277232010.3390/molecules27072320Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i>Guang-Ming Mei0Xiao-Hua Wu1Xiao-Jun Zhang2Jie Gu3Yi Fang4Chun-Ying Meng5Wen-Ge Yang6Zhejiang Provincial Key Lab of Mariculture & Enhancement, Zhejiang Marine Fisheries Research Institute, Tiyu Road 28, Zhoushan 316021, ChinaZhoushan Hospital Affiliated to Wenzhou Medical College, Dingshen Road 739, Zhoushan 316004, ChinaZhejiang Provincial Key Lab of Mariculture & Enhancement, Zhejiang Marine Fisheries Research Institute, Tiyu Road 28, Zhoushan 316021, ChinaZhejiang Provincial Key Lab of Mariculture & Enhancement, Zhejiang Marine Fisheries Research Institute, Tiyu Road 28, Zhoushan 316021, ChinaZhejiang Provincial Key Lab of Mariculture & Enhancement, Zhejiang Marine Fisheries Research Institute, Tiyu Road 28, Zhoushan 316021, ChinaZhejiang Provincial Key Lab of Mariculture & Enhancement, Zhejiang Marine Fisheries Research Institute, Tiyu Road 28, Zhoushan 316021, ChinaCollege of Food and Pharmaceutical Sciences, Ningbo University, Qixing South Road 169, Ningbo 315800, ChinaWe report here the purification of a novel metal-binding protein from <i>Oratosquilla oratoria</i> (<i>O. oratoria</i> MT-1) by gel and ion-exchange chromatography. SDS-PAGE and MALDI-TOF analyses demonstrated that isolated <i>O. oratoria</i> MT-1 was of high purity with a molecular weight of 12.4 kDa. The fluorescence response to SBD-F derivatives revealed that <i>O. oratoria</i> MT-1 contained a large number of sulfhydryl groups, which is a general property of metallothioneins. Zn and Cu metal stoichiometries for <i>O. oratoria</i> MT-1 were 3.97:1 and 0.55:1, respectively. The proportion of cysteine (Cys) residues in the amino acid composition was 32.69%, and aromatic amino acids were absent. The peptide sequence coverage with <i>Macrobrachium rosenbergii</i> calmodulin (accession AOA3S8FSK5) was 60%. Infrared spectroscopy of <i>O. oratoria</i> MT-1 revealed two obvious peaks at absorption frequencies for the amide I band and the amide II band. CD spectra revealed that the secondary structure was mainly composed of random coil (57.6%) and β-sheet (39.9%). An evaluation of in vitro antioxidant activity revealed that isolated <i>O. oratoria</i> MT-1 has strong reducing activities, exhibiting scavenging rates for DPPH and OH of 77.8% and 75.8%, respectively (IC<sub>50</sub> values 0.57 mg/mL and 1.1 mg/mL). <i>O. oratoria</i> MT-1 may be used as a functional additive in cosmetics, health foods, and medical products, as well as a reference material for quantitative analysis of metallothionein in such products.https://www.mdpi.com/1420-3049/27/7/2320<i>Oratosquilla oratoria</i>metallothioneinextractionpurificationantioxidant activity |
| spellingShingle | Guang-Ming Mei Xiao-Hua Wu Xiao-Jun Zhang Jie Gu Yi Fang Chun-Ying Meng Wen-Ge Yang Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i> Molecules <i>Oratosquilla oratoria</i> metallothionein extraction purification antioxidant activity |
| title | Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i> |
| title_full | Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i> |
| title_fullStr | Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i> |
| title_full_unstemmed | Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i> |
| title_short | Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from <i>Oratosquilla oratoria</i> |
| title_sort | structural characterization and in vitro antioxidant activity of metallothionein from i oratosquilla oratoria i |
| topic | <i>Oratosquilla oratoria</i> metallothionein extraction purification antioxidant activity |
| url | https://www.mdpi.com/1420-3049/27/7/2320 |
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