Direct desaturation of free myristic acid by hen liver microsomal Delta9-desaturase without prior activation to myristoyl-CoA derivative.
Direct desaturation of free myristic acid by hen liver microsomal Delta(9)-desaturase without prior activation to myristoyl-CoA by the addition of adenosine triphosphate (ATP) and CoA was observed when the incubation medium was mixed at mixing speeds of >250 rpm in the presence of fatty acid-binding proteins (FABP). Desaturation was linear with time and proportional to the microsomal protein concentration. Desaturation was maximal at pH 7.9. The greatest desaturation rate was observed at a mixing speed of 500 rpm in the presence of FABP. Desaturation decreased at mixing speeds of >500 rpm. Data suggest that when myristic acid is bound to FABP in the form of protein-monomer complexes, its activation to the CoA derivative is not necessary for it to be desaturated by the Delta(9)-desaturase when using mixing rates of >250 rpm. Myristic acid-FABP complexes serve as substrates for the Delta(9)-desaturase at mixing rates of >250 rpm. Desaturation was reduced by bovine serum albumin and alpha-bromohexadecanoate, and no desaturation was observed in the absence of FABP. These findings suggest that FABP may regulate the accessibility of fatty acids in the desaturation reaction to the active site of the desaturase rather than just protect the membrane-bound desaturase from the cytotoxic effect of free fatty acids.
Awad
Aziz C
AC
Department of Food Science and Human Nutrition, Michigan State University, East Lansing, Michigan 48824-1224, USA.
Shin
Han-Seung
HS
Romsos
Dale R
DR
Gray
J Ian
JI
D000214:Acyl Coenzyme A / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000494:pharmacology; D000818:Animals; D002352:Carrier Proteins / Q000378:metabolism; D002645:Chickens; D003065:Coenzyme A / Q000494:pharmacology; D050556:Fatty Acid-Binding Proteins; D006863:Hydrogen-Ion Concentration; D008862:Microsomes, Liver / Q000201:enzymology*; D019814:Myristic Acid / Q000378:metabolism*; D009243:NAD / Q000378:metabolism; D013230:Stearoyl-CoA Desaturase / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature
D000214:Acyl Coenzyme A; D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D019814:Myristic Acid; D009243:NAD; C040349:S-tetradecanoyl-coenzyme A; D000255:Adenosine Triphosphate; D013230:Stearoyl-CoA Desaturase; D003065:Coenzyme A