fa78685e9c0b908903b4727144687500 ijms_a2017v18n7p1457.pdf b5b8dbee3b028fd8cc70bfbdf17f8ef1c6182f35 ijms_a2017v18n7p1457.pdf a89bb12028b7d18383a46d0a3fefe50be107266a49d0fb50164e35974dd55e07 ijms_a2017v18n7p1457.pdf Title: Biomphalaria glabrata Metallothionein: Lacking Metal Specificity of the Protein and Missing Gene Upregulation Suggest Metal Sequestration by Exchange Instead of through Selective Binding Subject: The wild-type metallothionein (MT) of the freshwater snail Biomphalaria glabrata and a natural allelic mutant of it in which a lysine residue was replaced by an asparagine residue, were recombinantly expressed and analyzed for their metal-binding features with respect to Cd2+, Zn2+ and Cu+, applying spectroscopic and mass-spectrometric methods. In addition, the upregulation of the Biomphalaria glabrata MT gene was assessed by quantitative real-time detection PCR. The two recombinant proteins revealed to be very similar in most of their metal binding features. They lacked a clear metal-binding preference for any of the three metal ions assayed—which, to this degree, is clearly unprecedented in the world of Gastropoda MTs. There were, however, slight differences in copper-binding abilities between the two allelic variants. Overall, the missing metal specificity of the two recombinant MTs goes hand in hand with lacking upregulation of the respective MT gene. This suggests that in vivo, the Biomphalaria glabrata MT may be more important for metal replacement reactions through a constitutively abundant form, rather than for metal sequestration by high binding specificity. There are indications that the MT of Biomphalaria glabrata may share its unspecific features with MTs from other freshwater snails of the Hygrophila family. Keywords: metallothionein; metal-binding; zinc; copper; cadmium; Biomphalaria glabrata; Gastropoda; Hygrophila Author: Michael Niederwanger, Sara Calatayud, Oliver Zerbe, Sílvia Atrian, Ricard Albalat, Mercè Capdevila, Òscar Palacios and Reinhard Dallinger Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.15 CreationDate: Thu Jul 6 11:27:27 2017 CEST ModDate: Thu Jul 6 11:27:27 2017 CEST 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: 2394512 bytes Optimized: no PDF version: 1.5 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- GRSSNH+URWPalladioL-Ital Type 1 Custom yes yes no 109 0 ZQLYMX+URWPalladioL-BoldItal Type 1 Custom yes yes no 110 0 FMQRTR+URWPalladioL-Bold Type 1 Custom yes yes no 111 0 CTYSVZ+VnURWPalladioL-Bold Type 1 Custom yes yes no 112 0 HDBXJB+URWPalladioL-Roma Type 1 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