Evidence for the involvement of a 66 kDa membrane protein in the synthesis of sterolglucoside in Saccharomyces cerevisiae

• Autorzy: Lenart U. , Haplova J. , Magdolen P. , Farkas V. , Palamarczyk G.
• Języki publikacji: EN

Abstrakty

EN

The membrane-bound sterolglucoside synthase from the yeast Saccharomyces cerevisiae has been solubilized by nonionic detergent, Nonidet P-40, Triton X-100, and partially purified by DEAE-cellulose column chromatography and ammonium sulfate fractionation. SDS/PAGE of the purified fraction revealed the presence of two protein bands of molecular mass 66 kDa and 54 kDa. In an attempt to identify further the polypeptide chain of sterolglucoside synthase, the partially purified enzyme was treated with [di-125I]-5-[3-(p-azidosalicylamide)]allyl-UDPglucose, a photoactive analogue of UDPglucose, which is a substrate for this enzyme. Upon photolysis the 12SI-labeled probe was shown to link covalently to the 66kDa protein. The photoinsertion was competed out by the presence of unlabeled UDPglucose thus suggesting that this protein contains substrate binding site for UDPglucose. Since photoinsertion of the probe to protein of 66 kDa correlates with the molecular mass of the protein visualized upon enzyme purification we postulate that the 66 kDa protein is involved in sterolglucoside synthesis in yeast.

Słowa kluczowe

EN

yeast; Triton X-100; glucosylphosphodolichol; sterolglucoside synthesis; dithiothreitol; photoaffinity labeling; nonionic detergent; Nonidet P-40; phenylmethylsulphonyl fluoride; Saccharomyces cerevisiae; membrane protein

• Wydawca: -
• Czasopismo: Acta Biochimica Polonica
• Rocznik: 1995
• Tom: 42
• Numer: 2
• Opis fizyczny: p.269-274,fig.

Twórcy

autor

Lenart U.

• Polish Academy of Sciences, A.Pawinskiego 5a, 02-106 Warsaw, Poland

autor

Haplova J.

autor

Magdolen P.

autor

Farkas V.

autor

Palamarczyk G.

Bibliografia

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