e1e7a67c94779a528fa3d4bf0d2cfbb9 ijms_22_13114_v2.pdf 3feb19863f1058edb1db67bc34f7613f87c5d8c7 ijms_22_13114_v2.pdf 0c1ad9d91c8a4b89cb5cd68df423859ee3cc76b450da275cf36f637c62c25838 ijms_22_13114_v2.pdf Title: Metal-Specificity Divergence between Metallothioneins of Nerita peloronta (Neritimorpha, Gastropoda) Sets the Starting Point for a Novel Chemical MT Classification Proposal Subject: Metallothioneins’ (MTs) biological function has been a matter of debate since their discovery. The importance to categorize these cysteine-rich proteins with high coordinating capacity into a specific group led to numerous classification proposals. We proposed a classification based on their metal-binding abilities, gradually sorting them from those with high selectivity towards Zn/Cd to those that are Cu-specific. However, the study of the NpeMT1 and NpeMT2isoforms of Nerita peloronta, has put a new perspective on this classification. N. peloronta has been chosen as a representative mollusk to elucidate the metal-binding abilities of Neritimorpha MTs, an order without any MTs characterized recently. Both isoforms have been recombinantly synthesized in cultures supplemented with ZnII, CdII, or CuII, and the purified metal–MT complexes have been thoroughly characterized by spectroscopic and spectrometric methods, leading to results that confirmed that Neritimorpha share Cd-selective MTs with Caenogastropoda and Heterobranchia, solving a so far unresolved question. NpeMTs show high coordinating preferences towards divalent metal ions, although one of them (NpeMT1) shares features with the so-called genuine Zn-thioneins, while the other (NpeMT2) exhibits a higher preference for Cd. The dissimilarities between the two isoforms let a window open to a new proposal of chemical MT classification. Keywords: metallothioneins; Cd-selective; Neritimorpha; biochemical classification Author: Mario García-Risco, Sara Calatayud, Veronika Pedrini-Martha, Ricard Albalat, Reinhard Dallinger, Òscar Palacios and Mercè Capdevila Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Dec 6 03:31:07 2021 CET ModDate: Mon Dec 6 03:38:54 2021 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2112418 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- NLOJMS+VnURWPalladioL Type 1 Custom yes yes no 10 0 ZQJDAD+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 LNDCBS+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 FXDZMG+URWPalladioL-BoldItal Type 1 Custom yes yes no 24 0 ZPEQKZ+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 CTYSVZ+VnURWPalladioL-Bold Type 1 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261 0 FDLELO+Arial CID TrueType Identity-H yes yes yes 264 0 FDLAIB+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 280 0 FDLAID+PalatinoLinotype TrueType WinAnsi yes yes no 283 0 FDLAIF+PalatinoLinotype,Italic TrueType WinAnsi yes yes no 286 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-05-27 02:50:32 CEST RepresentationInformation: ijms_22_13114_v2.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 12:08:24 CEST Size: 2112418 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 513 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@44c8afef Item: Title: Results and Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Heterologous Expression and Production of Metal-NpeMT Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: The Metal-Binding Abilities of NpeMT Isoforms towards Divalent Metal Ions Confirms a Cd-Selective Origin in Snail MTs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Cd Selectivity in NpeMTs Makes Way to a Gradual Transition towards Zn Specificity Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: NpeMT Isoforms Reveal Important Differences in Their CuI Binding Features Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: NpeMT Isoforms Provide the Key to Set Up a Novel MT Classification Scheme Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Biological Implications Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Children: Item: Title: Cloning, Production, and Purification of Recombinant Metal MT Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Characterization of the Metal-NpeMT1 and Metal-NpeMT2 Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Metal-Protein Binding Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Info: Title: Metal-Specificity Divergence between Metallothioneins of Nerita peloronta (Neritimorpha, Gastropoda) Sets the Starting Point for a Novel Chemical MT Classification Proposal Author: Mario García-Risco, Sara Calatayud, Veronika Pedrini-Martha, Ricard Albalat, Reinhard Dallinger, Òscar Palacios and Mercè Capdevila Subject: Metallothioneins’ (MTs) biological function has been a matter of debate since their discovery. The importance to categorize these cysteine-rich proteins with high coordinating capacity into a specific group led to numerous classification proposals. We proposed a classification based on their metal-binding abilities, gradually sorting them from those with high selectivity towards Zn/Cd to those that are Cu-specific. However, the study of the NpeMT1 and NpeMT2isoforms of Nerita peloronta, has put a new perspective on this classification. N. peloronta has been chosen as a representative mollusk to elucidate the metal-binding abilities of Neritimorpha MTs, an order without any MTs characterized recently. Both isoforms have been recombinantly synthesized in cultures supplemented with ZnII, CdII, or CuII, and the purified metal MT complexes have been thoroughly characterized by spectroscopic and spectrometric methods, leading to results that confirmed that Neritimorpha share Cd-selective MTs with Caenogastropoda and Heterobranchia, solving a so far unresolved question. NpeMTs show high coordinating preferences towards divalent metal ions, although one of them (NpeMT1) shares features with the so-called genuine Zn-thioneins, while the other (NpeMT2) exhibits a higher preference for Cd. The dissimilarities between the two isoforms let a window open to a new proposal of chemical MT classification. 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