Cloning and Expression of a Seed-Specific Metallothionein-Like Protein from Sesame

Chyan, Chia-Lin; Lee, Tiger T. T.; Liu, Chiung-Pin; Yang, Yuan-Chang; Tzen, Jason T.C.; Chou, Wing‐Ming

doi:10.1271/bbb.69.2319

Cited by 45 publications

(22 citation statements)

References 32 publications

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“…The 14-kDa protein is probably a dimer, which is common in SDS-PAGE electrophoresis (Chatterjee and Maiti, 1987;Carpene et al, 2007). However, more possible reason might be that the retarded migration caused by the status of MTs binding metal ions (Chyan et al, 2005). The similar molecular weight of MT-like proteins also have been reported in other fish, e.g., the channel catfish (Ictalurus punctatus) (Chatterjee and Maiti, 1987), killfish (Heterandria formosa) (Xie and Klerks, 2004) and the mollusk, for example, the zebra mussel (Dressena polymorpha) (High et al, 1997).…”

Section: Discussionmentioning

confidence: 99%

Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization

Gao

Wang

Chen

et al. 2009

Comparative Biochemistry and Physiology Part C: Toxicology &

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Section: Discussionmentioning

confidence: 99%

Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization

Gao

Wang

Chen

et al. 2009

Comparative Biochemistry and Physiology Part C: Toxicology &

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“…However, among others the NMR structures clearly show that wheat E c -1 has the ability to coordinate six Zn II ions, and that the metal ion content is slightly lower than predicted due to the presence of a ZnCys 2 His 2 site, i.e., a site with a metal ion-to-ligand ratio of 1:4. In light of this, a content of 2.4 Zn II ions as determined for Sesamum indicum (sesame) clearly falls out of the range [39].…”

Section: >> Insertmentioning

confidence: 92%

Structural features specific to plant metallothioneins

Freisinger

2011

J Biol Inorg Chem

109

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The metallothionein (MT) superfamily combines a large variety of small cysteine-rich proteins from nearly all phyla of life that have the ability to coordinate various transition metal ions, including Zn(II), Cd(II), and Cu(I). The members of the plant MT family are characterized by great sequence diversity, requiring further subdivision into four subfamilies. Very peculiar and not well understood is the presence of rather long cysteine-free amino acid linkers between the cysteine-rich regions. In light of the distinct differences in sequence to MTs from other families, it seems obvious to assume that these differences will also be manifested on the structural level. This was already impressively demonstrated with the elucidation of the three-dimensional structure of the wheat E(c)-1 MT, which revealed two metal cluster arrangements previously unprecedented for any MT. However, as this structure is so far the only one available for the plant MT family, other sources of information are in high demand. In this review the focus is thus set on any structural features known, deduced, or assumed for the plant MT proteins. This includes the determination of secondary structural elements by circular dichroism, IR, and Raman spectroscopy, the analysis of the influence of the long linker regions, and the evaluation of the spatial arrangement of the sequence separated cysteine-rich regions with the aid of, e.g., limited proteolytic digestion. In addition, special attention is paid to the contents of divalent metal ions as the metal ion to cysteine ratios are important for predicting and understanding possible metal-thiolate cluster structures. Very peculiar and not well understood is the presence of rather long Cys-free amino acid linkers between the Cys-rich regions. In light of the distinct differences in sequence to MTs from other families it seems obvious to assume that these differences will be also manifested on the structural level. This was already impressively demonstrated with the elucidation of the three-dimensional structure of the wheat E c -1 MT, which revealed two metal cluster arrangements previously unprecedented for any MT. However, as this structure is so far the only one available for the plant MT family other sources of information are of high demand.In this review the focus is thus set on any structural features known, deduced or assumed for the plant MT proteins. This includes the determination of secondary structural elements by CD, IR, and Raman spectroscopy, the analysis of the influence of the long linker regions, as well as the evaluation of the spatial arrangement of the sequence separated Cys-rich regions with the aid of e.g. limited proteolytic digestion. In addition, special attention is paid to the contents of divalent metal ions as the metal ion-to-Cys ratios are important for predicting and

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“…MT4 did not respond to additional metal supplementation of Zn, Cu, or Cd. Previous results revealed that recombinant MT4 from sesame (Sesamum indicum; SiMT4) purified and subjected to elemental analysis by inductively coupled plasma atomic emission spectroscopy behaved similarly to barley MT4, as SiMT4 was found to coordinate two Zn ions (Chyan et al, 2005). However, extensive studies on both native and recombinant wheat Ec protein have shown that this protein has the capacity to coordinate up to six Zn ions (Leszczyszyn et al, 2007;Peroza and Freisinger, 2007).…”

Section: Differential Metal Speciation By Mt3 and Mt4mentioning

confidence: 99%

Barley Metallothioneins: MT3 and MT4 Are Localized in the Grain Aleurone Layer and Show Differential Zinc Binding

et al. 2012

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Metallothioneins (MTs) are low-molecular-weight, cysteine-rich proteins believed to play a role in cytosolic zinc (Zn) and copper (Cu) homeostasis. However, evidence for the functional properties of MTs has been hampered by methodological problems in the isolation and characterization of the proteins. Here, we document that barley (Hordeum vulgare) MT3 and MT4 proteins exist in planta and that they differ in tissue localization as well as in metal coordination chemistry. Combined transcriptional and histological analyses showed temporal and spatial correlations between transcript levels and protein abundance during grain development. MT3 was present in tissues of both maternal and filial origin throughout grain filling. In contrast, MT4 was confined to the embryo and aleurone layer, where it appeared during tissue specialization and remained until maturity. Using state-of-the-art speciation analysis by size-exclusion chromatography inductively coupled plasma mass spectrometry and electrospray ionization time-of-flight mass spectrometry on recombinant MT3 and MT4, their specificity and capacity for metal ion binding were quantified, showing a strong preferential Zn binding relative to Cu and cadmium (Cd) in MT4, which was not the case for MT3. When complementary DNAs from barley MTs were expressed in Cu-or Cd-sensitive yeast mutants, MT3 provided a much stronger complementation than did MT4. We conclude that MT3 may play a housekeeping role in metal homeostasis, while MT4 may function in Zn storage in developing and mature grains. The localization of MT4 and its discrimination against Cd make it an ideal candidate for future biofortification strategies directed toward increasing food and feed Zn concentrations.

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Cloning and Expression of a Seed-Specific Metallothionein-Like Protein from Sesame

Cited by 45 publications

References 32 publications

Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization

Metallothionein-2 gene from the mandarin fish Siniperca chuatsi: cDNA cloning, tissue expression, and immunohistochemical localization

Structural features specific to plant metallothioneins

Barley Metallothioneins: MT3 and MT4 Are Localized in the Grain Aleurone Layer and Show Differential Zinc Binding

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