Release of highly active Fet3 from membranes of the yeast Pichia pastoris by limited proteolysis.
Arch Biochem Biophys
; 372(2): 295-9, 1999 Dec 15.
Article
in En
| MEDLINE
| ID: mdl-10600167
ABSTRACT
A soluble derivative of Fet3 has been obtained from the methylotrophic yeast Pichia pastoris by limited proteolysis of membrane suspensions with trypsin. The soluble protein and the membrane-bound parent Fet3 have been purified to apparent homogeneity. Soluble Fet3 had molecular mass 100 kDa, while the full-length protein had molecular mass 110 kDa, in line with the expected decrease for cleavage and loss of a single transmembrane helix and a small cytoplasmic domain. The optical and EPR spectra of Fet3 were typical of the multicopper oxidases, indicating the presence of one type 1 blue copper site and a type 2/type 3 copper trinuclear cluster. V(max) values for iron oxidation by P. pastoris Fet3 were obtained similar to human ceruloplasmin and much higher than those reported for Saccharomyces cerevisiae Fet3.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Endopeptidases
/
Pichia
/
Ceruloplasmin
/
Cell Membrane
Limits:
Humans
Language:
En
Journal:
Arch Biochem Biophys
Year:
1999
Document type:
Article
Affiliation country:
Italy