Carnitine fatty acid transport

Production of Malonyl-CoA for the Fatty Acid Biosynthesis. Acetyl-CoA serves as a substrate in the production of malonyl-CoA. There are several routes by which acetyl-CoA is supplied to die cytoplasm. One route is the transfer of acetyl residues from the mitochondrial matrix across the mitochondrial membrane into the cyto-plasm. This process resembles a fatty acid transport and is likewise effected with the participation of carnitine and the enzyme acetyl-CoA-camitine transferase. Another route is the production of acetyl-CoA from citrate. Citrate is delivered from the mitochondria and undergoes cleavage in the cytoplasm by the action of the enzyme ATP-citrate lyase ... 

In 1955, Fritz determined that carnitine plays an essential role in fatty acid -oxidation (FAO), and in 1973 the first two clinically relevant disorders affecting this pathway were described primary carnitine deficiency by Engel and Angelini, and carnitine palmitoyltransferase (CPT) type II (CPT-II) deficiency by DiMauro and DiMauro [6, 7]. To date, more than 20 different enzyme deficiency states affecting fatty acid transport and mitochondrial / -oxidaLion have been described [8] and additional enzymes involved in this pathway are still being discovered [9, 10]. 

The study reported in Table 4.12 illustrates the role of carnitine in fatty add oxidation and introduces the topic of medium-chain fatty acids. This study, conducted before the role of carnitine in fatty acid transport was realized, involved addition of radioactive fatly acids to suspensions of liver mitochondria. The fatty acids used iriduded [ "Cjoctanoic acid (medium-chain) and [ " Clpalmitic add (long-chain). The susperisions were incubated for 30 minutes to permit uptake of the fatty acids, their subsequent oxidation, and discharge of radioactive carbon dioxide. The produced in the Krebs cycle diffuses out of the mitochondria into the surrounding fluid-... 

Fatty acids are more efficiently degraded because of increased synthesis of molecules that facilitate fatty acid transport and oxidation. The concentration of citric acid cycle and (3-oxidation enzymes increases, as well as the components of the ETC. In addition, the capacity of the muscle cell to remove fatty acids from blood and to transport them into mitochondria increases. For example, increases in the synthesis of fatty acid transporter proteins and fatty acid-binding proteins, as well as carnitine and carnitine acyltransferase, have been observed. 

Fatty acid transport proteins The mitochondrial and peroxisomal enzymes of fatty acid oxidation Carnitine palmitoyl transferase I HMG-CoA synthase Apoprotein Clll (suppression)... 

How are fatty acids transported to the mitochondrion for oxidation After an initial activation step in the cytosol, with formation of an acyl-CoA corresponding to each fetty acid, each acyl group is transesterified to carnitine for transport across the intermembrane space of the mitochondrion. The acyl group is again transesterified to form an acyl-GoA. 

This CoA derivative then exchanges its CoA for another partner called carnitine. The carnitine-fatty acid complex, which is soluble in the mitochondrial membranes, is then transported from the cytosol past the inner mitochondrial membrane into the inner matrix, where it exchanges the carnitine for acetyl-CoA, to again become the fatty acyl-CoA derivative. 

Although the balance between glucose and fatty acid oxidation is described in Chap. 13, it is relevant to note here that malonyl-CoA inhibits carnitine acyl transferase I (CAT-1), the enzyme that catalyzes the exchange of fatty acids for carnitine as part of the cytosol-to-matrix fatty acid transport system. Inhibition of CAT-I occurs when acetyl-CoA carboxylase is activated by insulin. By inhibiting the uptake of fatty acids into mitochondria, malonyl CoA favors the oxidation of glucose and prevents fatty acids from being oxidized at the same time as they are being synthesized. 

The mitochondrial outer membrane enz5mie carnitine palmitoyltransferase 1 (CPTl) is a main site of regulation of intracellular long-chain fatty acid transport. At least two isoforms of CPTl are expressed in the body L-CPTl (the liver-type isoform) and M-CPTl (the muscle-type isoform). Skin fibroblasts from healthy humans are known to contain only one isoform of CPTl the liver-type, which is encoded by the gene CPTIA. Skin fibroblasts from patients with a liver-type CPTl deficiency do not express either of the two known CPTl isoforms (neither liver- nor muscle-t3T)e), and therefore could provide an excellent background to study CPTl by means of molecular complementation. 

The carnitine fatty acid esters, which are in equilibrium with long chain acyl-CoA molecules in living muscle tissue, are of biochemical importance. The carnitine fatty acid ester, but not the acyl-CoA ester, can traverse the inner mitochondrial membrane. After the fatty acid is oxidized within the mitochondria, carnitine is instrumental in transporting the generated acetic acid out of the mitochondria. 

Ascorbic acid is involved in carnitine biosynthesis. Carnitine (y-amino-P-hydroxybutyric acid, trimethylbetaine) (30) is a component of heart muscle, skeletal tissue, Uver and other tissues. It is involved in the transport of fatty acids into mitochondria, where they are oxidized to provide energy for the ceU and animal. It is synthesized in animals from lysine and methionine by two hydroxylases, both containing ferrous iron and L-ascorbic acid. Ascorbic acid donates electrons to the enzymes involved in the metabohsm of L-tyrosine, cholesterol, and histamine (128). 

All of the other enzymes of the /3-oxidation pathway are located in the mitochondrial matrix. Short-chain fatty acids, as already mentioned, are transported into the matrix as free acids and form the acyl-CoA derivatives there. However, long-chain fatty acyl-CoA derivatives cannot be transported into the matrix directly. These long-chain derivatives must first be converted to acylearnitine derivatives, as shown in Figure 24.9. Carnitine acyltransferase I, located on the outer side of the inner mitochondrial membrane, catalyzes the formation of... 

Figure 22-1. Role of carnitine in the transport of long-chain fatty acids through the inner mitochondrial membrane. Long-chain acyl-CoA cannot pass through the inner mitochondrial membrane, but its metabolic product, acylcarnitine, can. 

Hepatic steatosis usually is a result of excessive administration of carbohydrates and/or lipids, but deficiencies of carnitine, choline, and essential fatty acids also may contribute. Hepatic steatosis can be minimized or reversed by avoiding overfeeding, especially from dextrose and lipids.35,38 Carnitine is an important amine that transports long-chain triglycerides into the mitochondria for oxidation, but carnitine deficiency in adults is extremely rare and is mostly a problem in premature infants and patients receiving chronic dialysis. Choline is an essential amine required for synthesis of cell membrane components such as phospholipids. Although a true choline deficiency is rare, preliminary studies of choline supplementation to adult patients PN caused reversal of steatosis. 

Long-chain fatty acids can slowly cross the mitochondrial membrane by themselves, but this is too slow to keep up with their metabolism. The carnitine shuttle provides a transport mechanism and allows control of (3 oxidation. Malonyl-CoA, a precursor for fatty acid synthesis, inhibits the carnitine shuttle and slows down (3 oxidation (Fig. 13-5). 

The CARNITINE SHUTTLE is used to transport fatty acids into the mitochondria. 

Carnitine is a vitamin-like quaternary ammonium salt, playing an important role in the human energy metabolism by facilitating the transport of long-chained fatty acids across the mitochondrial membranes. An easy, fast, and convenient procedure for the separation of the enantiomers of carnitine and 0-acylcarnitines has been reported on a lab-made teicoplanin-containing CSP [61]. The enantioresolution of carnitine and acetyl carnitine was enhanced when tested on a TAG CSP, prepared in an identical way [45]. Higher a values were reached also in the case of A-40,926 CSP [41]. 

Carnitine acyltransfefase-I (CAT-1) and carnitine acyltran erase-2 (CAT-2) are also refened to as carnitine palmrtoyl transferase-1 (CPT-1) and carnitine palmitoyl transferase-2 (CPT-2). The carnitine transport system is most important for allowing long-chain fatty acids to enter into the mitochondria. 

Long-chain fatty acids must be activated and transported into the mitochondria. Fatty acyl CoA synthetase, on the outer mitochondrial membrane, activates the fatty adds by attaching CoA. The fetty acyl portion is then transferred onto carnitine by carnitine aqdtransferase-I for transport into the mitochondria. The sequence of events is shown in Figure 1-16-2 and indudes the following steps ... 

Mitochondria contain all the enzymes necessary for oxidation of fatty acids but, before this can take place, the fatty acids have to be transported into the mitochondria. Transport requires the formation of an ester of the fatty acid with a compound, carnitine, in a reaction catalysed by the enzyme carnitine palmitoyltransferase ... 

Figure 7.11 Mechanism of transport of long-chain fatty adds across the inner mitochondrial membrane as fatty acyl carnitine. CRT is the abbreviation for carnitine palmitoyl transferase. CPT-I resides on the outer surface of the inner membrane, whereas CPT-II resides on the inner side of the inner membrane of the mitochondria. Transport across the inner membrane is achieved by a carrier protein known as a translocase. FACN - fatty acyl carnitine, CN - carnitine. Despite the name, CRT reacts with long-chain fatty acids other than palmitate. CN is transported out of the mitochondria by the same translocase.

The most important process in the degradation of fatty acids is p-oxidation—a metabolic pathway in the mitochondrial matrix (see p. 164). initially, the fatty acids in the cytoplasm are activated by binding to coenzyme A into acyl CoA [3]. Then, with the help of a transport system (the carnitine shuttle [4] see p. 164), the activated fatty acids enter the mitochondrial matrix, where they are broken down into acetyl CoA. The resulting acetyl residues can be oxidized to CO2 in the tricarboxylic acid cycle, producing reduced... 

After uptake by the cell, fatty acids are activated by conversion into their CoA derivatives—acyl CoA is formed. This uses up two energy-rich anhydride bonds of ATP per fatty acid (see p. 162). For channeling into the mitochondria, the acyl residues are first transferred to carnitine and then transported across the inner membrane as acyl carnitine (see B). 

The inner mitochondrial membrane has a group-specific transport system for fatty acids. In the cytoplasm, the acyl groups of activated fatty acids are transferred to carnitine by carnitine acyltransferase [1 ]. They are then channeled into the matrix by an acylcar-nitine/carnitine antiport as acyl carnitine, in exchange for free carnitine. In the matrix, the mitochondrial enzyme carnitine acyltransferase catalyzes the return transfer of the acyl residue to CoA. 

Fatty acids with an odd number of C atoms are treated in the same way as normal fatty acids—i. e., they are taken up by the cell with ATP-dependent activation to acyl CoA and are transported into the mitochondria with the help of the carnitine shuttle and broken down there by p-oxidation (see p. 164). In the last step, propionyl CoA arises instead of acetyl CoA. This is first carboxylated by propionyl CoA carboxylase into fSj-methylmalonyl CoA [3], which—after isomerization into the (i ) enantiomer (not shown see p. 411)—is isomerized into succinyl CoA [4]. 

C. Long-chain fatty acids (LCFAs), which have carbon chain lengths of 12-22 units (C12-C22), must be transported into the mitochondrial matrix where the enzymes responsible for their oxidation are located. This is accomplished by the carnitine shutde (Figure 8-3). 

The enzymes of fatty acid oxidation in animal cells are located in the mitochondrial matrix, as demonstrated in 1948 by Eugene P. Kennedy and Albert Lehninger. The fatty acids with chain lengths of 12 or fewer carbons enter mitochondria without the help of membrane transporters. Those with 14 or more carbons, which constitute the majority of the FFA obtained in the diet or released from adipose tissue, cannot pass directly through the mitochondrial membranes—they must first undergo the three enzymatic reactions of the carnitine shuttle. The first reaction is catalyzed by a family of isozymes (different isozymes specific for fatty acids having short, intermediate, or long carbon chains) present... 

FIGURE17-6 Fatty acid entry into mitochondria via the acyl-carnitine/ carnitine transporter. After fatty acyl-carnitine is formed at the outer membrane or in the intermembrane space, it moves into the matrix by facilitated diffusion through the transporter in the inner membrane. In the matrix, the acyl group istransferred to mitochondrial coenzyme... 

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