Role of external loops of human ceruloplasmin in copper loading by ATP7B and Ccc2p.
Ceruloplasmin is a multicopper oxidase required for correct iron homeostasis.Previously, we have identified a ceruloplasmin mutant associated with the iron overload disease aceruloplasminemia, which was unable to acquire copper from the mammalian pump ATP7B but could be produced in an enzymatically active form in yeast. Here, we report the expression of recombinant ceruloplasmin in the yeast Pichia pastoris and the study of the role of five surface-exposed loops in copper incorporation by comparing the efficiencies of mammalian ATP7B and yeast Ccc2p. The possibility to "mix and match" mammalian and yeast multicopper oxidases and copper ATPases can provide clues on the molecular features underlying the process of copper loading in multicopper oxidases.
Maio
Nunziata
N
Dipartimento di Biologia, Università ROMA TRE, 00146 Rome.
Polticelli
Fabio
F
De Francesco
Giovanni
G
Rizzo
Gianluca
G
Bonaccorsi di Patti
Maria Carmela
MC
Musci
Giovanni
G
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000818:Animals; D001665:Binding Sites; D027682:Cation Transport Proteins / Q000378:metabolism*; D002570:Ceruloplasmin / Q000737:chemistry / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D003300:Copper / Q000378:metabolism / Q000494:pharmacology*; D000073840:Copper-Transporting ATPases; D005816:Genetic Complementation Test; D006801:Humans; D008322:Mammals; D008958:Models, Molecular; D010843:Pichia / Q000378:metabolism; D011487:Protein Conformation; D020033:Protein Isoforms / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000187:drug effects / Q000378:metabolism; D014714:Vertebrates
D027682:Cation Transport Proteins; D020033:Protein Isoforms; D011994:Recombinant Proteins; D003300:Copper; D002570:Ceruloplasmin; D000251:Adenosine Triphosphatases; C000612464:ATP7B protein, human; D000073840:Copper-Transporting ATPases