Assay of acyl-CoA:monoglyceride acyltransferase from rat small intestine using continuous recording spectrophotometry.
Acyl-CoA:monoglyceride acyltransferase in microsomal preparations from the small intestine of the rat has been measured by means of continuous recording spectrophotometry. The reaction of 5,5'-dithiobis(2-nitrobenzoic acid) with CoA has been employed for this assay and optimal conditions for the reaction have been defined. One of the substrates, palmitoyl-CoA, inhibits the reaction even in modest concentrations. This inhibition is largely prevented by the addition of bovine serum albumin to the incubation medium. The reliability of the assay method was confirmed by comparison with the more cumbersome assay method that uses radioactive substrate.
D000217:Acyltransferases / Q000032:analysis* / Q000037:antagonists & inhibitors; D000818:Animals; D001565:Benzoates; D002417:Cattle; D003065:Coenzyme A; D003720:Densitometry; D003902:Detergents; D005246:Feedback; D005989:Glycerides; D006863:Hydrogen-Ion Concentration; D007413:Intestinal Mucosa / Q000201:enzymology; D007421:Intestine, Small / Q000201:enzymology*; D008297:Male; D008722:Methods; D008861:Microsomes / Q000201:enzymology*; D009578:Nitrobenzenes; D009829:Oleic Acids; D010169:Palmitic Acids; D051381:Rats; D012710:Serum Albumin, Bovine; D013053:Spectrophotometry
D001565:Benzoates; D003902:Detergents; D005989:Glycerides; D009578:Nitrobenzenes; D009829:Oleic Acids; D010169:Palmitic Acids; D012710:Serum Albumin, Bovine; D000217:Acyltransferases; D003065:Coenzyme A