Purification and characterization of a novel specific phosphatidylglycerol-phosphatidylinositol transfer protein with high activity from Aspergillus oryzae.
A novel phospholipid transfer protein has been purified to homogeneity 406-fold from the filamentous fungus Aspergillus oryzae. The successive steps of purification comprised ultrafiltration, gel filtration on Sephadex G-75, ion exchange chromatographies on DEAE-Sepharose and Mono Q. The active protein is a monomer with a molecular mass of 19,000, estimated from SDS electrophoresis, amino acid composition as well as gel filtration. The isoelectric point is 4.8. The amino acid composition is characterized by a high amount of Gly, Leu, Ser, Asx and Glx residues and 4 Cys residues. N-terminal sequence was determined and compared with M. mucedo sequence. The purified protein was found to transfer preferentially phosphatidylglycerol and phosphatidylinositol over phosphatidylcholine > phosphatidylethanolamine > phosphatidylserine and no phosphatidic acid. Optimal temperature for in vitro transfer was 25-30 degrees C and optimal pH 4-7. Heating protein at 100 degrees C does not inactivate protein whereas a denaturation with urea is irreversible.
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Laboratoire de Biotechnologie des Champignons Filamenteux, Faculté des Sciences de Luminy, Parc Scientifique et Technologique, Marseille, France.
Asther
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Marion
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Asther
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D000595:Amino Acid Sequence; D000818:Animals; D001236:Aspergillus oryzae / Q000737:chemistry*; D002352:Carrier Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002417:Cattle; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004355:Drug Stability; D004492:Edetic Acid / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007526:Isoelectric Point; D008565:Membrane Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010715:Phosphatidylglycerols / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism*; D049294:Phospholipid Transfer Proteins; D051381:Rats; D017385:Sequence Homology; D013696:Temperature; D014508:Urea / Q000494:pharmacology
D002352:Carrier Proteins; D008565:Membrane Proteins; D010715:Phosphatidylglycerols; D010716:Phosphatidylinositols; D049294:Phospholipid Transfer Proteins; D014508:Urea; D004492:Edetic Acid