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Enzymatic hydrolysis of sphingolipids

The acyl composition of sphingolipids is determined by the specificity of the enzyme ceramide synthase (EC 2.3.1.24). Ceramide contains a long-chain base (LCB) linked to a variety of fatty acids to produce a lipid class with potentially hundreds of structural variants. An optimized procedure for the assay of ceramide synthase in yeast microsomes is reported that uses mass spectrometry to detect any possible LCB and fatty acid combination synthesized from unlabeled substrates provided in the reaction. The assay requires the delivery of substrates with bovine serum albumin for maximum activity within defined limits of substrate concentration and specific methods to stop the reaction and extract the lipid that avoid the non-enzymatic synthesis of ceramide. The activity of ceramide synthase in yeast microsomes is demonstrated with the four natural LCBs found in yeast along with six saturated and two unsaturated fatty acyl-coenzyme As from 16 to 26 carbons in length. The procedure allows for the determination of substrate specificity and kinetic parameters toward natural substrates for ceramide synthase from potentially any organism.

First suggestion that sphingolipid-derived metabolitescould function as second messengers.

Cholesterol biosynthesis
80% of endogenous cholesterol is formed in the liver. Enzymes involved in the synthesis are partly located in the endoplasmic reticulum and partly in the cytoplasm. Acetyl CoA is the precursor. HMG CoA, mevalonate, isopentenyl pyrophosphate, squalene are some of the important intermediate compounds formed during cholesterol synthesis. Cholesterol is transported in lipoprotein complexes. Elevation of lipids in blood leads to the deposition of cholesterol on the arterial walls leading to atherosclerosis.

Enzymatic hydrolysis of sphingolipids V

Sphingolipid Metabolic Pathway: An Overview of Major …

Fatty acid oxidation
Fatty acids have to be activated prior to their entry into mitochondrial matrix where the enzymes of β-oxidation of fatty acids are located. Activated fatty acids are then transported from cytosol to the mitochondrial matrix with the help of carnitine transporter. Total net yield of ATP per molecule of palmitic acid is 129. Similarly oxidation of unsaturated and odd chain fatty acids also take place with additional reactions. Β-oxidation in peroxisomes involves three enzymatic reactions. Minor pathways of oxidation such as α-oxidation of branched chain fatty acids and ω-oxidation of medium and long chain fatty acids in microsomes do take place in our body.

Ketone body metabolism
Ketone bodies are acetoacetate, β-hydroxy butyrate and acetone. Ketone bodies are synthesized in the liver but they are utilized by extra hepatic tissues as fuels. Ketone bodies are accumulated in the blood if the rate of synthesis exceeds the ability of extra hepatic tissues to utilize them. This leads to excess ketone bodies in blood, excretion of ketone bodies in urine and smell of acetone in breath. All these three together are known as ketosis. In uncontrolled diabetes mellitus and starvation, ketone bodies are formed.

In view of the above considerations, what part can ozone play as an antiviral agent? In one study,[46] polio virus 1 was exposed to 0.21 mg/liter of ozone at pH 7.2. After 30 seconds 99% of the viruses were inactivated (lost their ability to replicate within host cells), but appeared to maintain their structural integrity. Analysis of viral components showed damage to polypeptide chains and envelope proteins, which could result in attachment capability compromise, and breakage of the single-stranded RNA into two parts, producing replicating dysfunction at its root level. Other researchers[54] in similar experiments concluded that in ozonation, it is the viral capsid which sustains damage. It is to be noted however, that the polioviridae (Picornavirus family) contain four structural proteins encapsulating a single RNA strand and are devoid of lipids.

Enzymatic Hydrolysis of Sphingolipids. VI. Hydrolysis …

there is an upregulation of the enzymes involved in the synthesis of complex sphingolipids

These phytosphingosine-containing lipids were the first sphingolipids to be described with structural features of a glycolipid and of a phospholipid.

Morton RA discovered and named ubiquinone (coenzyme Q) in the unsaponifiable fraction of liver lipids ()

Bernhard K et al.

Lipids are digested and absorbed with the help of bile salts. Products of lipid digestion aggregate to form mixed micelles and are absorbed into the small intestine. Lipids are transported in the form of lipoproteins. Fatty acids are activated, transported across mitochondrial membrane with the help of carnitine transporter. β -oxidation of saturated fatty acids takes place in the mitochondrial matrix. Similarly oxidation of unsaturated and odd chain fatty acids also take place with additional reactions. Ketone bodies are formed in the liver but they are utilized by extra hepatic tissues. In uncontrolled diabetes mellitus and starvation, excessive ketone bodies are formed, leading to ketosis. Fatty acid biosynthesis takes place in the cytosol of cells. Fat gets deposited in the adipose tissue. Acetyl Coenzyme A is the precursor of fatty acid synthesis as well as cholesterol biosynthesis. Elevation of lipids in blood leads to deposition of cholesterol plaques in the arterial walls leading to atherosclerosis. Prostaglandins and leukotrienes are synthesized from twenty carbon unsaturated fatty acids. Phosphatidic acid is an important intermediate in the synthesis of glycerophospholipids. In sphingolipids, sphingosine is present as an alcohol.

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Epidermal sphingolipids: Metabolism, function, and …

Synthesis of phospholipids and sphingolipids
Glycerol is the starting material in the synthesis of glycerophospholipids. CDP-diacylglycerol is the activated intermediate of this pathway. Sphingolipids are synthesized from palmitoyl CoA and serine, initially to form ceramide. Sphingomyelins and glycolipids are synthesized from ceramide. Various sphingolipids storage diseases are resulted due to hereditary absence of hydrolytic enzymes.

Epidermal sphingolipids: Metabolism, function, and roles ..

By means of several enzymaticreactions, cholesterol is synthesized from acetic acid; it then servesas the major precursor for the synthesis of vitamin D3, of the varioussteroid hormones, including cortisol, cortisone, and aldosterone in theadrenal glands, and of the sex hormones progesterone, estrogen, and testosterone.

Sphingolipid metabolism and cell signaling. Part A …

AB - The acyl composition of sphingolipids is determined by the specificity of the enzyme ceramide synthase (EC 2.3.1.24). Ceramide contains a long-chain base (LCB) linked to a variety of fatty acids to produce a lipid class with potentially hundreds of structural variants. An optimized procedure for the assay of ceramide synthase in yeast microsomes is reported that uses mass spectrometry to detect any possible LCB and fatty acid combination synthesized from unlabeled substrates provided in the reaction. The assay requires the delivery of substrates with bovine serum albumin for maximum activity within defined limits of substrate concentration and specific methods to stop the reaction and extract the lipid that avoid the non-enzymatic synthesis of ceramide. The activity of ceramide synthase in yeast microsomes is demonstrated with the four natural LCBs found in yeast along with six saturated and two unsaturated fatty acyl-coenzyme As from 16 to 26 carbons in length. The procedure allows for the determination of substrate specificity and kinetic parameters toward natural substrates for ceramide synthase from potentially any organism.

Sphingolipid metabolism and cell signaling

At the Golgi, ceramide takes part in a metabolic flux towards sphingomyelin, and glycosphingolipids, which drives lipid raft formation and towards the plasma membrane.

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