Fatty acyl-Co A: sphingosine acyltransferase in bovine brain mitochondria: its solubilization and reconstitution onto the membrane lipid liposomes

Biological & Pharmaceutical Bulletin
H ShimenoA Nagamatsu

Abstract

Fatty acyl-Co A: sphingosine acyltransferase (ceramide synthase, EC 2.3.1.24) is mainly localized in the microsomal and mitochondrial membranes. Attempts to isolate the enzyme have failed, largely because there has been little or no detection of the enzyme activity in detergent extracts. In this study, we solubilized the membrane-bound enzyme from bovine brain mitochondria with a Tris-HCl buffer containing 2% Triton X-100 and, after removal of the detergent, reconstituted it with the membrane lipid liposomes. The specific activity of the reconstituted enzyme was approx. 8 times higher than that of the solubilized enzyme. We next examined the lipid dependence of the enzyme, using various phospholipid liposomes. The ability of phospholipids to enhance the activity of solubilized ceramide synthase was specific and structure-related. The most potent stimulator was phosphatidylserine liposomes, suggesting an important role of the net negative charges. This paper also describes a highly reproducible high-performance liquid chromatographic (HPLC) procedure for the determination of ceramide synthase activity. Combination of the HPLC method with the reconstituted enzyme system appears to be suitable for elucidating the characteristics o...Continue Reading

Citations

Feb 28, 2009·Journal of Cardiovascular Pharmacology·Sergei A Novgorodov, Tatyana I Gudz
Dec 15, 2010·The Journal of Biological Chemistry·Sergei A NovgorodovTatyana I Gudz
Jul 18, 2001·Pathology Oncology Research : POR·I Peták, J A Houghton
Sep 18, 2002·The Journal of Cell Biology·Hervé Le StunffSarah Spiegel
Feb 25, 2010·FEBS Letters·Branka Stancevic, Richard Kolesnick
Jan 18, 2012·The Biochemical Journal·Thomas D MullenLina M Obeid
Apr 26, 2005·The Journal of Biological Chemistry·Jimmy A RotoloRichard Kolesnick
Aug 23, 2003·Oncogene·Heike GrassméErich Gulbins
Aug 30, 2003·Oncogene·Richard Kolesnick, Zvi Fuks
Feb 27, 2015·Biological Chemistry·Carla HajjAdriana Haimovitz-Friedman
Apr 4, 2018·Journal of Lipids·Xingxuan He, Edward H Schuchman
Dec 31, 2020·Journal of the American Society of Nephrology : JASN·Yelena DrexlerSandra Merscher
Feb 19, 2000·Cellular Signalling·R T Dobrowsky
Jun 13, 1998·Progress in Lipid Research·L RiboniG Tettamanti

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.