Reference: Bonaccorsi di Patti MC, et al. (1999) Release of highly active Fet3 from membranes of the yeast Pichia pastoris by limited proteolysis. Arch Biochem Biophys 372(2):295-9

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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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Journal Article | Research Support, Non-U.S. Gov't
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Bonaccorsi di Patti MC, Bellenchi GC, Bielli P, Calabrese L
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