Characterization of purified and unidirectionally reconstituted Pho84 phosphate permease of Saccharomyces cerevisiae

Citation
U. Fristedt et al., Characterization of purified and unidirectionally reconstituted Pho84 phosphate permease of Saccharomyces cerevisiae, FEBS LETTER, 458(1), 1999, pp. 1-5
Citations number
19
Categorie Soggetti
Biochemistry & Biophysics
Journal title
FEBS LETTERS
ISSN journal
00145793 → ACNP
Volume
458
Issue
1
Year of publication
1999
Pages
1 - 5
Database
ISI
SICI code
0014-5793(19990910)458:1<1:COPAUR>2.0.ZU;2-A
Abstract
Hydropathy analysis of the amino acid sequence of the Pho84 phosphate perme ase of Saccharomyces cerevisiae suggests that the protein consists of 12 tr ansmembrane domains connected by hydrophilic loops. The Pho84 protein has b een modified by a gene fusion approach, yielding two different N-terminal H is-tagged chimeras which can be expressed in Escherichia coli, purified and functionally reconstituted into defined proteoliposomes, The continuous ep itopes in the N- and C-terminal sequences of the Pho84 chimeras were shown to be accessible in proteoliposomes containing the purified active Pho84 pr oteins. Site-specific proteolysis of the immunoreactive N-terminal sequence in the reconstituted protein suggests a unidirectional insertion into lipo somes. (C) 1999 Federation of European Biochemical Societies.