Peroxisomal localization of acyl-coenzyme A reductase (long chain alcohol forming) in guinea pig intestine mucosal cells

• Published in: The Journal of biological chemistry 1991-07, Vol.266 (19), p.12201-12206
• Main Authors: BURDETT, K, LARKINS, L. K, DAS, A. K, HAJRA, A. K.
• Format: Article
• Language: English
• Subjects: Acyl-CoA Dehydrogenase, Long-Chain - metabolism; Analytical, structural and metabolic biochemistry; Animals; Biological and medical sciences; Cell Fractionation; Enzymes and enzyme inhibitors; fatty acyl-CoA: NADPH oxidoreductase; Fundamental and applied biological sciences. Psychology; Guinea Pigs; Intestinal Mucosa - cytology; Intestinal Mucosa - enzymology; Lipids - biosynthesis; Lysosomes - enzymology; Microbodies - enzymology; Mitochondria - enzymology; Oxidoreductases; peroxisomes; Trypsin - chemistry
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/s0021-9258(18)98881-2

Upon differential centrifugation of guinea pig intestine mucosal cells homogenate, fatty acyl-CoA:NADPH oxidoreductase (long chain alcohol forming) was found to be enriched in the light mitochondrial (L) fraction (sedimenting between 66,000 x g min and 500,000 x g min) which contained mainly mitochondria, lysosomes, and peroxisomes. Peroxisomes (marker enzymes: catalase and dihydroxyacetone phosphate acyltransferase) present in the L fraction were separated from other organelles in a Nycodenz density gradient centrifugation employing a vertical rotor. By comparing the distribution of acyl-CoA reductase with different marker enzymes in the gradient, it was concluded that this reductase is primarily localized in the microperoxisomes (microbodies). The topography of the membrane-bound enzyme in the isolated organelles was studied by checking its lability toward trypsin in the absence and presence of the detergent Triton X-100. The results suggested that acyl-CoA reductase is localized on the outer surface (cytosolic side) of microperoxisomal membrane.