Analysis of spectrometry and thermodynamics of the metallothionein in freshwater crab Sinopotamon henanense for its binding ability with different metals

Yang, Hui Zhen; Wang, Lu; He, Yong; Jing, Wei X.; Wen, Liang; Chen, Chien M; Wang, Lan

doi:10.1016/j.chemosphere.2019.125670

Cited by 6 publications

(1 citation statement)

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“…The Zn stoichiometry (approximately 4:1) of O. oratoria MT-1 was lower than that of MT-M-I and MT-M-II from Arca subcrenata (6:1 and 7:1) [ 33 ] and lower than that of an MT isoform from Homarus americanus , which exhibits a coordinating stoichiometry of six Zn 2+ or Cd 2+ [ 46 ]. The heterologous expressed MT of Sinopotamon henanense was observed to bind 15 equivalents of Zn 2+ in vivo (as analyzed by ESI-TOF-MS), but only six equivalents of Zn 2+ were required to form the stable metal-S coordination bonds in the tetrahedral structure that coordinates Zn [ 47 ]. In most cases, MTs bind 7–12 metal ions [ 48 ].…”

Section: Resultsmentioning

confidence: 99%

Structural Characterization and In Vitro Antioxidant Activity of Metallothionein from Oratosquilla oratoria

Mei

Zhang

et al. 2022

Molecules

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We report here the purification of a novel metal-binding protein from Oratosquilla oratoria (O. oratoria MT-1) by gel and ion-exchange chromatography. SDS-PAGE and MALDI-TOF analyses demonstrated that isolated O. oratoria MT-1 was of high purity with a molecular weight of 12.4 kDa. The fluorescence response to SBD-F derivatives revealed that O. oratoria MT-1 contained a large number of sulfhydryl groups, which is a general property of metallothioneins. Zn and Cu metal stoichiometries for O. oratoria 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 Macrobrachium rosenbergii calmodulin (accession AOA3S8FSK5) was 60%. Infrared spectroscopy of O. oratoria 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 O. oratoria MT-1 has strong reducing activities, exhibiting scavenging rates for DPPH and OH of 77.8% and 75.8%, respectively (IC50 values 0.57 mg/mL and 1.1 mg/mL). O. oratoria 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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