Enzymes acyl carnitine transferase

Carnitine serves as a cofactor for several enzymes, including carnitine translo-case and acyl carnitine transferases I and II, which are essential for the movement of activated long-chain fatty acids from the cytoplasm into the mitochondria (Figure 11.2). The translocation of fatty acids (FAs) is critical for the genaation of adenosine triphosphate (ATP) within skeletal muscle, via 3-oxidation. These activated FAs become esterified to acylcamitines with carnitine via camitine-acyl-transferase I (CAT I) in the outer mitochondrial membrane. Acylcamitines can easily permeate the membrane of the mitochondria and are translocated across the membrane by carnitine translocase. Carnitine s actions are not yet complete because the mitochondrion has two membranes to cross thus, through the action of CAT II, the acylcar-nitines are converted back to acyl-CoA and carnitine. Acyl-CoA can be used to generate ATP via 3-oxidation, Krebs cycle, and the electron transport chain. Carnitine is recycled to the cytoplasm for fumre use. 

The transport is accomplished with the participation of carnitine, which takes up the acyl from acyl-CoA on the outer membrane side. Acylcamitine assisted by carnitine translocase diffuses to the inner side of the membrane to give its acyl to the CoA located in the matrix. The process of reversible acyl transfer between CoA and carnitine on the outer and inner sides of the membrane is effected by the enzyme acyl-CoA-camitine transferase. 

The answer is b. (Murray, pp 505-626. Scriver, pp 4029-4240. Sack, pp 121-138. Wilson, pp 287-320.) A deficiency in carnitine, carnitine acyl-transferase 1, carnitine acyltransferase 11, or acylcarnitine translocase can lead to an inability to oxidize long-chain fatty acids. This occurs because all of these components are needed to translocate activated long-chain (>10 carbons long) fatty acyl CoA across mitochondrial inner membrane into the matrix where P oxidation takes place. Once long-chain fatty acids are coupled to the sulfur atom of CoA on the outer mitochondrial membrane, they can be transferred to carnitine by the enzyme carnitine acyltransferase I, which is located on the cytosolic side of the inner mitochondrial membrane. Acyl carnitine is transferred across the inner membrane to the matrix surface by translocase. At this point the acyl group is reattached to a CoA sulfhydryl by the carnitine acyltransferase 11 located on the matrix face of the inner mitochondrial membrane. 

CoASH and thus reforming the true immediate substrate for the oxidatioa According to this hypothesis, acyl carnitine, which is not a substrate for the enzymes of the oxidative process, is an admirable substrate for fatty acid degradation in intact mitochondrial systems. In addition, mitochondria depleted of their endogenous energy donors (ATP, GTP) are unable to oxidize add fatty acids unless ATP and carnitine are both present in the system. This observation clearly proves that acyl-CoA is formed outside of the inner mitochondrial membrane—i.e. outside of the oxidation compartment—and is transported to the inner compartment via the carnitine-linked transport mechanism. In agreement with these results an ATP-dependent thiokinase has been identified in the outer mitochondrial membrane, and an acyl-camitine transferase has been found in the inner mitochondrial membrane. 

At this point, the acyl-CoA is still in the cytosol of the muscle cell. Entry of the acyl-CoA into the mitochondrial matrix requires two translocase enzymes, carnitine acyl transferase I and carnitine acyl transferase II (CAT I and CAT II), and a carrier molecule called carnitine the carnitine shuttles between the two membranes. The process of transporting fatty acyl-CoA into mitochondria is shown in Figure 7.15. 

One enzyme regulated by AMPK is acetyl-CoA carboxylase, which produces malonyl-CoA, the first intermediate committed to fatty acid synthesis. Malonyl-CoA is a powerful inhibitor of the enzyme carnitine acyl-transferase I, which starts the process of ]3 oxidation by transporting fatty acids into the mitochondrion (see Fig. 17-6). By phosphorylating and inactivating acetyl-CoA carboxylase, AMPK inhibits fatty acid synthesis while relieving the inhibition (by malonyl-CoA) of )3 oxidation (Fig. 23-37). 

Carnitine acyltransferase 1 is strongly inhibited by malonyl CoA, andmuscle has both acetyl CoA carboxylase, which forms malonyl CoA, and malonyl CoA decarboxylase, which acts to remove malonyl CoA and relieve the inhibition of carnitine acyl transferase. The two enzymes are regulated in opposite directions in response to insulin, which stimulates fatty acid synthesis and reduces /S-oxidation, and glucagon that reduces fatty acid synthesis and increases p-oxidation (Kerner and Hoppel, 2000 Louet et al., 2001 Eaton, 2002). 

Bieber LL, Farel SS. Carnitine acyl transferases. Enzymes 1983 16 624-644. 

This hepatomegaly is correlated to PPARa activation of acyl-CoA oxidase (AOX), the first enzyme of peroxisomal /3-oxidation of fatty acids and a gene with a PPRE in its promoter region14. In addition, hepatic mitochondrial /3-oxidation and microsomal w-oxidation of fatty acids are increased, as a direct result of PPARa activation of mRNA of specific enzymes associated with these pathways (carnitine palmitoyl transferase I and cytochrome P4504A, respectively). Activation of fatty acid oxidation by these three pathways would lead to enhanced fatty acid oxidation, given the appropriate substrate. PPARa has also been shown to enhance delivery of fatty acids to the oxidizing systems (Fig. 3). 

Carnitine palmitoyltransferase I (CPTI also called carnitine acyltransferase I, CATI), the enzyme that transfers long-chain fatty acyl groups from CoA to carnitine, is located on the outer mitochondrial membrane (Fig. 23.5). Fatty acylcamitine crosses the inner mitochondrial membrane with the aid of a translocase. The fatty acyl group is transferred back to CoA by a second enzyme, carnitine palmitoyl-transferase II (CPTII or CATII). The carnitine released in this reaction returns to the cytosolic side of the mitochondrial membrane by the same translocase that brings fatty acylcamitine to the matrix side. Long-chain fatty acyl CoA, now located within the mitochondrial matrix, is a substrate for (3-oxidation. 

Fatty acid uptake by muscle requires the participation of fatty acid-binding proteins and the usual enzymes of fatty acid oxidation. Fatty acyl-CoA uptake into the mitochondria is controlled by malonyl-CoA, which is produced by an isozyme of acetyl-coA carboxylase (ACC-2 the ACC-1 isozyme is found in liver and adipose tissue and is used for fatty acid biosynthesis). ACC-2 is inhibited by phosphorylation by the AMP-activated protein kinase (AMP-PK) such that when energy levels are low the levels of malonyl CoA will drop, allowing fatty acid oxidation by the mitochondria. In addition, muscle cells also contain the enzyme malonyl CoA decarboxylase, which is activated by phosphorylation by the AMP-PK. Malonyl CoA decarboxylase converts malonyl CoA to acetyl CoA, thereby relieving the inhibition of carnitine palmitoyl transferase I (CPT-I) and stimulating fatty acid oxidation (Fig. 47.5). Muscle cells do not synthesize fatty acids the presence of acetyl CoA carboxylase in muscle is exclusively for regulatory purposes. 

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. 

In these experiments cultured glioma cells were exposed to TTA, and key enzymes in peroxisomal (fatty acyl-CoA oxidase—FAO) and mitochondrial (carnitine palmitoyl transferase II—CPT II) fatty acid oxidation were studied. 

Mitochondrial P-oxidation of long-chain fatty acids is the major source of energy production in man. The mitochondrial inner membrane is impermeable to long chain fatty acids or their CoA esters whereas acylcamitines are transported. Three different gene products are involved in this carnitine dependent transport shuttle carnitine palmi-toyl transferase I (CPT I), carnitine acyl-camitine carrier (CAC) and carnitine palmitoyl transferase II (CPT II). The first enzyme (CPT I) converts fatty acyl-CoA esters to their carnitine esters which are subsequently translocated across the mitochondrial inner membrane in exchange for free carnitine by the action of the carnitine acyl-camitine carrier (CAC). Once inside the mitochondrion, CPT II reconverts the carnitine ester back to the CoA ester which can then serve as a substrate for the P-oxidation spiral. 

As well as being the substrate for fatty acid synthesis, malonyl CoA has an important role in controlling (3-oxidation of fatty acids. Malonyl CoA is a potent inhibitor of carnitine palmitoyl transferase 1, the mitochondrial outer membrane enzyme that regulates uptake of fatty acyl CoA into the mitochondria (section 5.5.1). This means that, under conditions in which fatty acids are being synthesized in the cytosol, there will not be uptake into the mitochondria for (3-oxidation. (See also section 10.6.2.1 for a discussion of the role of malonyl CoA in regulating muscle fuel selection.)... 

Carnitine acyl transferase activity is controlled by malonyl CoA. As discussed in section 10.5.2, in liver and adipose tissue this serves to inhibit mitochondrial uptake and P-oxidation of fatty acids when fatty acids are being synthesized in the cytosol. Muscle also has an active acetyl CoA carboxylase, and synthesizes malonyl CoA, although it does not synthesize fatty acids, and muscle carnitine acyl transferase is more sensitive to inhibition by malonyl CoA than is the enzyme in liver and adipose tissue. 

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