Long-chain fatty acids activated to fatty acyl-CoA by acyl-CoA synthetase on the cytosolic face of endoplasmic reticulum and outer mitochondrial membrane cannot diffuse across the inner mitochondrial membrane, so a dedicated import shuttle is essential for matrix beta-oxidation. Carnitine palmitoyltransferase I, CPT I, localized to the outer membrane with catalytic site facing cytosol, transfers the acyl group to carnitine, forming acylcarnitine plus free CoA. Acylcarnitine is then shuttled across the inner membrane by the carnitine-acylcarnitine translocase, CACT, an antiporter exchanging acylcarnitine inward for free carnitine outward. At the matrix leaflet, carnitine palmitoyltransferase II, CPT II, reverses the reaction, regenerating fatty acyl-CoA and releasing carnitine for recycling. Inside the matrix, beta-oxidation spiral removes two carbons per cycle as acetyl-CoA through acyl-CoA dehydrogenase, enoyl-CoA hydratase, hydroxyacyl-CoA dehydrogenase and thiolase reactions, producing NADH and FADH2 that donate electrons to respiratory chain. Resulting acetyl-CoA feeds citric acid cycle for ATP synthesis. Malonyl-CoA produced during fed state allosterically inhibits CPT I, preventing futile cycling between synthesis and oxidation and coordinating metabolic switching.
Ref:
Berg et al., Biochemistry, 9th ed., Chapter 22: Fatty Acid Oxidation and Carnitine Shuttle.