Assay of acyl-CoA:monoglyceride acyltransferase from rat small intestine using continuous recording spectrophotometry

Abstract

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.

Keywords

• small bowel mucosa
• microsomal preparation
• substrate requirements
• substrate inhibition
• bovine serum albumin
• nonionic detergent