Rat liver outer mitochondrial carnitine palmitoyltransferase activity towards long-chain polyunsaturated fatty acids and their CoA esters.
The activity of the overt form of rat liver mitochondrial carnitine palmitoyltransferase or CPT0 (EC 2.3.1.21) towards different fatty acid substrates was studied. The following non-esterified fatty acids (NEFA) and their CoA esters in the presence of 1% bovine serum albumin (BSA) were tested: 16:0, 18:0, 18:1, 18:2, 18:3 omega 3, 20:4, 20:5 omega 3 and 22:6 omega 3. The data fit a square hyperbolic model for enzyme catalysis (p less than 0.001, non-linear regression). Asymptotic Vmax and K0.5, substrate concentration at one-half Vmax, were calculated using total concentrations of acyl-CoA, or unbound concentrations of NEFA. BSA was found to act as a true substrate reservoir for NEFA in that the dissociation of the NEFA-BSA complex was 10-330 times faster than the CPT0 reaction. Regardless of form (NEFA or CoA ester), 18:3 omega 3 gave the highest, while 22:6 omega 3 and 18:0 gave the lowest rates of acylcarnitine synthesis. Except for 18:3 omega 3 and 18:2, Vmax for NEFA was generally lower than for acyl-CoA, with the greatest differences observed for 20:4, 20:5 omega 3 and 22:6 omega 3, suggesting that acyl-CoA synthesis may also be important in the control of the entry of these fatty acids into the mitochondria. The data provide an enzymatic rationale for the relatively low content of 18:3 omega 3 in esterified lipid.
Gavino
G R
GR
Department of Nutrition, Université de Montreal, Quebec, Canada.
Gavino
V C
VC
D000214:Acyl Coenzyme A / Q000378:metabolism*; D000818:Animals; D002334:Carnitine O-Palmitoyltransferase / Q000378:metabolism*; D005230:Fatty Acids, Nonesterified / Q000032:analysis / Q000378:metabolism; D005231:Fatty Acids, Unsaturated / Q000378:metabolism*; D007425:Intracellular Membranes / Q000201:enzymology; D007700:Kinetics; D008297:Male; D008930:Mitochondria, Liver / Q000201:enzymology*; D011485:Protein Binding; D051381:Rats; D011919:Rats, Inbred Strains; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013367:Submitochondrial Particles / Q000201:enzymology; D013379:Substrate Specificity
D000214:Acyl Coenzyme A; D005230:Fatty Acids, Nonesterified; D005231:Fatty Acids, Unsaturated; D012710:Serum Albumin, Bovine; D002334:Carnitine O-Palmitoyltransferase