Effect of membrane perturbants on the activity and phase distribution of inositol phosphorylceramide synthase; development of a novel assay

Biochemistry
Paul A AeedAke P Elhammer

Abstract

The effect of 26 different membrane-perturbing agents on the activity and phase distribution of inositol phosphorylceramide synthase (IPC synthase) activity in crude Candida albicans membranes was investigated. The nonionic detergents Triton X-100, Nonidet P-40, Brij, Tween, and octylglucoside all inactivated the enzyme. However, at moderate concentrations, the activity of the Triton X-100- and octylglucoside-solubilized material could be partially restored by inclusion of 5 mM phosphatidylinositol (PI) in the solubilization buffer. The apparent molecular mass of IPC synthase activity solubilized in 2% Triton X-100 was between 1.5 x 10(6) and 20 x 10(6) Da, while under identical conditions, octylglucoside-solubilized activity remained associated with large presumably membrane-like structures. Increased detergent concentrations produced more drastic losses of enzymatic activity. The zwitterionic detergents Empigen BB, N-dodecyl-N,N-(dimethylammonio)butyrate (DDMAB), Zwittergent 3-10, and amidosulfobetaine (ASB)-16 all appeared capable of solubilizing IPC synthase. However, these agents also inactivated the enzyme essentially irreversibly. Solubilization with lysophospholipids again resulted in drastic losses of enzymatic activit...Continue Reading

References

Aug 9, 1996·The Journal of Biological Chemistry·Y LavieM C Cabot
Jan 24, 1998·The Journal of Biological Chemistry·S M MandalaM B Kurtz
Oct 6, 1998·Annual Review of Biochemistry·R C Dickson
Mar 20, 1999·The Journal of Biological Chemistry·S Strahl-Bolsinger, A Scheinost
Aug 1, 1959·Canadian Journal of Biochemistry and Physiology·E G BLIGH, W J DYER

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Citations

Dec 3, 2008·Antimicrobial Agents and Chemotherapy·Paul A AeedAke P Elhammer
Mar 20, 2014·The Journal of Organic Chemistry·Anna Alcaide, Amadeu Llebaria
Nov 16, 2007·Organic & Biomolecular Chemistry·David MormeneoAntonio Delgado
Oct 1, 2012·Drug Delivery and Translational Research·Byeong Hee HwangSamir Mitragotri
Jun 24, 2015·ACS Medicinal Chemistry Letters·Peter G M WutsAke P Elhammer
Jun 22, 2010·The International Journal of Biochemistry & Cell Biology·John G MinaPaul W Denny

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