Transfer of cholesteryl esters and phospholipids as well as net deposition by microsomal triglyceride transfer protein.

Journal of Lipid Research
Paul RavaM Mahmood Hussain

Abstract

Microsomal triglyceride transfer protein (MTP) activity is classically measured using radioactive lipids. We described a simple fluorescence assay to measure its triacylglycerol (TAG) transfer activity. Here, we describe fluorescence-based methods to measure the transfer of phospholipids (PLs) and cholesteryl esters (CEs) by MTP. Both transfer activities increased with time and MTP amounts and were inhibited to different extents by an MTP antagonist, BMS197636. We also describe a method to measure the net deposition of fluorescent lipids in acceptor vesicles. In this procedure, negatively charged donor vesicles are incubated with MTP and acceptor vesicles, and lipids transferred to acceptors are quantified after the removal of donor vesicles and MTP by the addition of DE52. Lipid deposition in acceptor vesicles was dependent on time and MTP. Using these methods, TAG transfer activity was the most robust activity present in purified MTP; CE and PL transfer activities were 60-71% and 5-13% of the TAG transfer activity, respectively. The method to determine lipid transfer is recommended for routine MTP activity measurements for its simplicity. These methods may help identify specific inhibitors for individual lipid transfer activi...Continue Reading

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Citations

May 27, 2008·The Journal of Biological Chemistry·Jahangir IqbalM Mahmood Hussain
Aug 10, 2005·The Journal of Experimental Medicine·Stephanie K DouganRichard S Blumberg
Feb 22, 2007·The Journal of Experimental Medicine·Stephanie K DouganRichard S Blumberg
Nov 26, 2009·The Journal of Pharmacology and Experimental Therapeutics·Yi LuoXian-Cheng Jiang
Sep 2, 2010·The Journal of Pharmacology and Experimental Therapeutics·Yi LuoXian-Cheng Jiang
Feb 23, 2012·Nutrition & Metabolism·M Mahmood HussainJahangir Iqbal
Sep 3, 2009·Journal of Lipid Research·Harmen WiersmaUwe J F Tietge
Jan 13, 2012·Journal of Lipid Research·Mathilde Di FilippoAgnès Sassolas
Nov 29, 2011·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·Irani KhatunM Mahmood Hussain
Mar 21, 2013·Hepatology Research : the Official Journal of the Japan Society of Hepatology·Xiang WangHiromi Rakugi
Feb 15, 2011·Biochemical and Biophysical Research Communications·Janet D SparksCharles E Sparks
Aug 9, 2011·Current Biology : CB·Nicholas DourisCarla B Green
May 8, 2008·Cell Metabolism·Jahangir IqbalM Mahmood Hussain
Oct 4, 2016·Critical Reviews in Clinical Laboratory Sciences·Meghan T Walsh, M Mahmood Hussain
Aug 9, 2006·Journal of Lipid Research·Sumbul FatmaM Mahmood Hussain
Jun 22, 2007·Journal of Lipid Research·Kamran AnwarM Mahmood Hussain
Apr 14, 2018·Journal of Lipid Research·Alaa Sirwi, M Mahmood Hussain
Apr 5, 2007·The Journal of Biological Chemistry·Xiaoyue PanM Mahmood Hussain
Jun 19, 2007·The Journal of Biological Chemistry·Xiaoyue Pan, M Mahmood Hussain

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