title: Ionic self-complementarity induces amyloid-like fibril formation in an isolated domain of a plant copper metallochaperone protein.
abstract: BACKGROUND
  Arabidopsis thaliana copper metallochaperone CCH is a functional homologue of yeast antioxidant ATX1, involved in cytosolic copper transport. In higher plants, CCH has to be transported to specialised cells through plasmodesmata, being the only metallochaperone reported to date that leaves the cell where it is synthesised. CCH has two different domains, the N-terminal domain conserved among other copper-metallochaperones and a C-terminal domain absent in all the identified non-plant metallochaperones. The aim of the present study was the biochemical and biophysical characterisation of the C-terminal domain of the copper metallochaperone CCH.
  
  
  RESULTS
  The conformational behaviour of the isolated C-domain in solution is complex and implies the adoption of mixed conformations in different environments. The ionic self-complementary peptide KTEAETKTEAKVDAKADVE, derived from the C-domain of CCH, adopts and extended conformation in solution with a high content in beta-sheet structure that induces a pH-dependent fibril formation. Freeze drying electron microscopy studies revealed the existence of well ordered amyloid-like fibrils in preparations from both the C-domain and its derivative peptide.
  
  
  CONCLUSIONS
  A number of proteins related with copper homeostasis have a high tendency to form fibrils. The determinants for fibril formation, as well as the possible physiological role are not fully understood. Here we show that the plant exclusive C-domain of the copper metallochaperone CCH has conformational plasticity and forms fibrils at defined experimental conditions. The putative influence of these properties with plant copper delivery will be addressed in the future.
authors.affiliation: Departament de Bioquímica i Biologia Molecular, Universitat de València, E-46100 Burjassot, València, Spain. helena.mira@uv.es
authors.forename: Helena
authors.initials: H
authors.lastname: Mira
authors.forename: Marçal
authors.initials: M
authors.lastname: Vilar
authors.forename: Vicent
authors.initials: V
authors.lastname: Esteve
authors.forename: Marc
authors.initials: M
authors.lastname: Martinell
authors.forename: Marcelo J
authors.initials: MJ
authors.lastname: Kogan
authors.forename: Ernest
authors.initials: E
authors.lastname: Giralt
authors.forename: David
authors.initials: D
authors.lastname: Salom
authors.forename: Ismael
authors.initials: I
authors.lastname: Mingarro
authors.forename: Lola
authors.initials: L
authors.lastname: Peñarrubia
authors.forename: Enrique
authors.initials: E
authors.lastname: Pérez-Payá
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D001692:Biological Transport / Q000502:physiology; D003300:Copper / Q000378:metabolism*; D018832:Molecular Chaperones / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D008969:Molecular Sequence Data; D009994:Osmolar Concentration; D010455:Peptides / Q000737:chemistry* / Q000378:metabolism / Q000502:physiology; D011487:Protein Conformation; D017433:Protein Structure, Secondary / Q000502:physiology; D000685:Serum Amyloid A Protein / Q000737:chemistry*
chemical_list: D029681:Arabidopsis Proteins; D018832:Molecular Chaperones; D010455:Peptides; D000685:Serum Amyloid A Protein; D003300:Copper