Release of highly active Fet3 from membranes of the yeast Pichia pastoris by limited proteolysis

Maria Carmela Bonaccorsi Di Patti, Gian Carlo Bellenchi, Pamela Bielli, Lilia Calabrese

Research output: Contribution to journalArticle

13 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)295-299
Number of pages5
JournalArchives of Biochemistry and Biophysics
Volume372
Issue number2
DOIs
Publication statusPublished - Dec 15 1999

Fingerprint

Proteolysis
Pichia
Molecular mass
Yeast
Copper
Yeasts
Membranes
Ceruloplasmin
Trypsin
Paramagnetic resonance
Saccharomyces cerevisiae
Suspensions
Oxidoreductases
Membrane Proteins
Iron
Derivatives
Oxidation
Proteins

Keywords

  • Ferroxidase activity
  • Fet3
  • Pichia pastoris

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Release of highly active Fet3 from membranes of the yeast Pichia pastoris by limited proteolysis. / Bonaccorsi Di Patti, Maria Carmela; Bellenchi, Gian Carlo; Bielli, Pamela; Calabrese, Lilia.

In: Archives of Biochemistry and Biophysics, Vol. 372, No. 2, 15.12.1999, p. 295-299.

Research output: Contribution to journalArticle

Bonaccorsi Di Patti, Maria Carmela ; Bellenchi, Gian Carlo ; Bielli, Pamela ; Calabrese, Lilia. / Release of highly active Fet3 from membranes of the yeast Pichia pastoris by limited proteolysis. In: Archives of Biochemistry and Biophysics. 1999 ; Vol. 372, No. 2. pp. 295-299.
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