Acyl-carnitine transferase

When large amounts of exogenous fatty acid enter the liver, how can they be transferred into the mitochondrion, for beta oxidation, with malonyl CoA inhibition This dichotomy is overcome because fatty-acyl CoAs inhibit acetyl CoA carboxylase and the malonyl CoA present proceeds onto fatty acids. When the malonyl CoA is converted to fatty acid, the level of malonyl CoA drops and is not restored. Thus, the inhibition of acyl carnitine transferase 1 is removed and fatty-acid oxidation can proceed. 

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. 

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.

Fig. 3.2.3 Profiles of acylcarnitines as their butyl esters in plasma (precursor of m/z 85 scan) of a normal control (a) and three patients with elevated Gi-acyl carnitine (m/z 288 peak 4) that represents primarily butyrylcarnitine in a patient with short-chain acyl-CoA dehydrogenase (SCAD) deficiency (b), isobutyrylcarnitine in a patient with isobutyryl-CoA dehydrogenase (IBDH) deficiency (c), and a natural isotope of formiminoglutamate (FIGLU m/z 287 peak 3) in a patient with glutamate formimino-transferase deficiency (d). Peak 1 free carnitine (m/z 218), peak 2 acetylcarnitine (C2 m/z 260). The asterisks represent the internal standards (from left to right) d3-acetylcarnitine (C2 m/z 263), d3-propionylcarnitine (C3 m/z 277), d3-butyrylcarnitine (C4 m/z 291), d3-octanoylcarnitine (C8 m/z 347), d3-dodecanoylcarnitine (Ci, m/z 403), and d3-pal-mitoylcarnitine (Ci6 m/z 459)...

Answer The transport of fatty acid molecules into mitochondria requires a shuttle system involving a fatty acyl-carnitine intermediate. Fatty acids are first converted to fatty acyl-CoA molecules in the cytosol (by the action of acyl-CoA synthetases) then, at the outer mitochondrial membrane, the fatty acyl group is transferred to carnitine (by the action of carnitine acyl-transferase I). After transport of fatty acyl-carnitine through the inner membrane, the fatty acyl group is transferred to mitochondrial CoA. The cytosolic and mitochondrial pools of CoA are thus kept separate, and no labeled CoA from the cytosolic pool enters the mitochondrion. 

But a transferase now comes upon the scene Making fatty acyl carnitine. 

Fatty acids are activated on the outer mitochondrial membrane, whereas they are oxidized in the mitochondrial matrix. A special transport mechanism is needed to carry long-chain acyl CoA molecules across the inner mitochondrial membrane. Activated long-chain fatty acids are transported across the membrane by conjugating them to carnitine, a zwitterionic alcohol. The acyl group is transferred from the sulfur atom of CoA to the hydroxyl group of carnitine to form acyl carnitine. This reaction is catalyzed by carnitine acyltransferase I (also called carnitine palmitoyl transferase I), which is bound to the outer mitochondrial membrane. 

A on the matrix side of the membrane. This reaction, which is catalyzed by carnitine acy[transferase II (carnitine palmitoyl transferase II), is simply the reverse of the reaction that takes place in the cytoplasm. The reaction is thermodynamically feasible because of the zwitterionic nature of carnitine The O-acyl link in carnitine has a high group-transfer potential, apparently because, being zwitterions, carnitine and its esters are solvated differently from most other alcohols and their esters. Finally, the translocase returns carnitine to the cytoplasmic side in exchange for an incoming acyl carnitine. 

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. 

The control of fatty-acid oxidation is related to the availability of circulating fatty acids and the activity of palmitoyl carnitine transferase 1. When circulating fatty acids are elevated, considerable fatty-acyl CoA is formed in a number of tissues, including the liver, which is sufficient to inhibit both acetyl CoA carboxylase in the cytosol and, indirectly, pyruvate dehydrogenase in the mitochondrion. Under this condition, neither malonyl CoA nor citrate would accumulate thus, there would be a diminution of fatty-acid synthesis. When large amounts of fatty... 

Fig. 23.5. Transport of long-chain fatty acids into mitochondria. The fatty acyl CoA crosses the outer mitochondrial membrane. Carnitine pahnitoyl transferase I in the outer mitochondrial membrane transfers the fatty acyl group to carnitine and releases Co ASH. The fatty acyl carnitine is translocated into the mitochondrial matrix as carnitine moves out. Carnitine palmitoyl transferase II on the inner mitochondrial membrane transfers the fatty acyl group back to CoASH, to form fatty acyl CoA in the matrix.

Within the peroxisome, the acetyl groups can be transferred from CoA to carnitine by an acetylcarnitine transferase, or they can enter the cytosol. A similar reaction converts medium-chain-length acyl CoAs and the short-chain butyryl CoA to acyl carnitine derivatives. These acylcarnitines diffuse from the peroxisome to the mitochondria, pass through the outer mitochondrial membrane, and are transported through the inner mitochondrial membrane via the carnitine translocase system. 

P-Oxidation of fatty acids is controlled by the uptake of fatty acids into the mitochondria - as discussed in section 5.5.1, this is controlled by the activity of carnitine acyl transferase on the outer mitochondrial membrane, and by the countertransport of acyl-carnitine and free carnitine across 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. 

The steps in the subsequent utilization of muscle LCFAs may be summarized as follows. The free fatty acids, liberated from triglycerides by a neutral triglyceride lipase, are activated to form acyl CoAs by the mediation of LCFAcyl-CoA synthetase which is situated on the outer mitochondrial membrane. The next step involves carnitine palmitoyl transferase I (CPT I, see Figure 9) which is also located on the outer mitochondrial membrane and catalyzes the transfer of LCFAcyl residues from CoA to carnitine (y-trimethyl-amino-P-hydroxybutyrate). LCFAcyl... 

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. 

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). 

Malonyl CoA is an allosteric effector of carnitine acyl transferase. What kind of effector is it, i.e., activator or inhibitor, and what is the logic behind the interaction ... 

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