5b8f0cb5879d391507a9f890b8c45d64 pmc_33374169.pdf 6433fa235b614836740e7acfdd8676c6b7e3e950 pmc_33374169.pdf 93808f56b4431e41563173c1f2ed87a50084f6c85a9dc5b620b22cc462749f25 pmc_33374169.pdf Title: Two Unconventional Metallothioneins in the Apple Snail Pomacea bridgesii Have Lost Their Metal Specificity during Adaptation to Freshwater Habitats Subject: Metallothioneins (MTs) are a diverse group of proteins responsible for the control of metal homeostasis and detoxification. To investigate the impact that environmental conditions might have had on the metal-binding abilities of these proteins, we have characterized the MTs from the apple snail Pomacea bridgesii, a gastropod species belonging to the class of Caenogastropoda with an amphibious lifestyle facing diverse situations of metal bioavailability. P. bridgesii has two structurally divergent MTs, named PbrMT1 and PbrMT2, that are longer than other gastropod MTs due to the presence of extra sequence motifs and metal-binding domains. We have characterized the Zn(II), Cd(II), and Cu(I) binding abilities of these two MTs after their heterologous expression in E. coli. Our results have revealed that despite their structural differences, both MTs share an unspecific metal-binding character, and a great ability to cope with elevated amounts of different metal ions. Our analyses have also revealed slight divergences in their metal-binding features: PbrMT1 shows a more pronounced Zn(II)-thionein character than PbrMT2, while the latter has a stronger Cu(I)-thionein character. The characterization of these two unconventional PbrMTs supports the loss of the metal-binding specificity during the evolution of the MTs of the Ampullariid family, and further suggests an evolutionary link of this loss with the adaptation of these gastropod lineages to metal-poor freshwater habitats. Keywords: metallothionein; metal-specificity; protein domains; Pomacea bridgesii; Mollusca; Gastropoda Author: Mario García-Risco, Sara Calatayud, Michael Niederwanger, Ricard Albalat, Òscar Palacios, Mercè Capdevila and Reinhard Dallinger Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Dec 24 09:17:39 2020 CET ModDate: Fri Dec 25 09:44:02 2020 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 15 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 1917451 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- JZJYCG+VnURWPalladioL Type 1 Custom yes yes no 10 0 GCJHBL+URWPalladioL-Ital Type 1 Custom yes yes no 15 0 HITBJT+URWPalladioL-Bold Type 1 Custom yes yes no 20 0 WNBUBO+URWPalladioL-BoldItal Type 1 Custom yes yes no 24 0 DVVALR+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 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Identity-H yes yes yes 151 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 157 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 163 0 FNTSBS+ArialMT CID TrueType Identity-H yes yes yes 169 0 FNTSBS+Calibri CID TrueType Identity-H yes yes yes 175 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 187 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 193 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 199 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 205 0 OEYAKW+Dingbats Type 1 Builtin yes yes no 217 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 223 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 229 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 235 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 241 0 FNTSBS+PalatinoLinotype-Italic CID TrueType Identity-H yes yes yes 258 0 FNTSBS+PalatinoLinotype-Bold CID TrueType Identity-H yes yes yes 264 0 FNTSBS+PalatinoLinotype-Roman CID TrueType Identity-H yes yes yes 270 0 FNTSBS+PalatinoLinotype-BoldItalic CID TrueType Identity-H yes yes yes 276 0 FNTSBS+TimesNewRomanPSMT CID TrueType Identity-H yes yes yes 282 0 Jhove (Rel. 1.22.1, 2019-04-17) Date: 2023-05-27 02:49:53 CEST RepresentationInformation: pmc_33374169.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2023-03-31 12:08:29 CEST Size: 1917451 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 499 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 Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7b69c6ba Children: Item: Title: Sequence Features of PbrMTs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@46daef40 Item: Title: Production of Metal PbrMT Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@12f41634 Item: Title: Zn(II) Binding Abilities of the PbrMT Isoforms Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Cd(II) Binding Abilities of the PbrMT Isoforms Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Item: Title: Cu(I) Binding Abilities of the PbrMTs Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Discussion 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 Pomacea bridgesii Metallothioneins for Heterologous Expression Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Production and Purification of Recombinant Metal PbrMT Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Characterization of the Metal PbrMT1 and Metal PbrMT2 Complexes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: In Vitro Metal Protein Binding Studies Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Info: Title: Two Unconventional Metallothioneins in the Apple Snail Pomacea bridgesii Have Lost Their Metal Specificity during Adaptation to Freshwater Habitats Author: Mario García-Risco, Sara Calatayud, Michael Niederwanger, Ricard Albalat, Òscar Palacios, Mercè Capdevila and Reinhard Dallinger Subject: Metallothioneins (MTs) are a diverse group of proteins responsible for the control of metal homeostasis and detoxification. To investigate the impact that environmental conditions might have had on the metal-binding abilities of these proteins, we have characterized the MTs from the apple snail Pomacea bridgesii, a gastropod species belonging to the class of Caenogastropoda with an amphibious lifestyle facing diverse situations of metal bioavailability. P. bridgesii has two structurally divergent MTs, named PbrMT1 and PbrMT2, that are longer than other gastropod MTs due to the presence of extra sequence motifs and metal-binding domains. We have characterized the Zn(II), Cd(II), and Cu(I) binding abilities of these two MTs after their heterologous expression in E. coli. Our results have revealed that despite their structural differences, both MTs share an unspecific metal-binding character, and a great ability to cope with elevated amounts of different metal ions. Our analyses have also revealed slight divergences in their metal-binding features: PbrMT1 shows a more pronounced Zn(II)-thionein character than PbrMT2, while the latter has a stronger Cu(I)-thionein character. The characterization of these two unconventional PbrMTs supports the loss of the metal-binding specificity during the evolution of the MTs of the Ampullariid family, and further suggests an evolutionary link of this loss with the adaptation of these gastropod lineages to metal-poor freshwater habitats. 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