Proteomic analysis of fructophilic properties of osmotolerant Candida magnoliae.
Candida magnoliae, an osmotolerant and erythritol producing yeast, prefers D-fructose to D-glucose as carbon sources. For the investigation of the fructophilic characteristics with respect to sugar transportation, a sequential extraction method using various detergents and ultracentrifugation was developed to isolate cellular membrane proteins in C. magnoliae. Immunoblot analysis with the Pma1 antibody and twodimensional electrophoresis analysis coupled with MS showed that the fraction II was enriched with membrane proteins. Eighteen proteins out of 36 spots were identified as membrane or membrane-associated proteins involved in sugar uptake, stress response, carbon metabolism, and so on. Among them, three proteins were significantly upregulated under the fructose supplying conditions. The hexose transporter was highly homologous to Ght6p in Schizosaccharomyces pombe, which was known as a predominant transporter for the fructose uptake of S. pombe because it exhibited higher affinity to D-fructose than D-glucose. The physicochemical properties of the ATP-binding cassette transporter and inorganic transporter explained their direct or indirect associations with the fructophilic behavior of C. magnoliae. The identification and characterization of membrane proteins involved in sugar uptake might contribute to the elucidation of the selective utilization of fructose to glucose by C. magnoliae at a molecular level.
Yu
Ji-Hee
JH
Department of Agricultural Biotechnology and the BIO-MAX Institute, Seoul National University, Seoul 151-921, Korea.
Lee
Dae-Hee
DH
Park
Yong-Cheol
YC
Lee
Mi-Gi
MG
Kim
Dae-Ok
DO
Ryu
Yeon-Woo
YW
Seo
Jin-Ho
JH
D018528:ATP-Binding Cassette Transporters / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D000595:Amino Acid Sequence; D002175:Candida / Q000737:chemistry* / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D005632:Fructose / Q000378:metabolism*; D005656:Fungal Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D005947:Glucose / Q000378:metabolism; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D008969:Molecular Sequence Data; D009004:Monosaccharide Transport Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009997:Osmotic Pressure; D028061:Phosphate Transport Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D040901:Proteomics; D016415:Sequence Alignment
D018528:ATP-Binding Cassette Transporters; D005656:Fungal Proteins; D008565:Membrane Proteins; D009004:Monosaccharide Transport Proteins; D028061:Phosphate Transport Proteins; D005632:Fructose; D005947:Glucose