Distribution of 3-hydroxy-3-methylglutaryl-CoA reductase in isolated villus and crypt cells of chick duodenum, jejunum and ileum.
3-Hydroxy-3-methylglutaryl-CoA reductase (EC 1.1.1.34), the major rate-limiting enzyme of cholesterogenesis, was studied in epithelial cells isolated in a villus to crypt gradient from chick duodenum, jejunum and ileum, in order to resolve the apparent controversy that exists on the anatomical localization of sterol synthesis in the intestine. Consistent separation was demonstrated by using the marker enzymes alkaline phosphatase, specific to the villus cells, and thymidine kinase, specific to the crypt cells. No relative difference in stability was observed, as shown by the equal distribution of acid phosphatase. Cells were 90-95 per cent viable. The highest specific activity of reductase was located in the microsomal fraction (41 per cent of the total). The mitochondria had lower specific activity (8 per cent of the total). The distribution of reductase activity in epithelial cells of the villus-crypt axis was also studied. The specific activity in each cell fraction from chick duodenum was clearly lower than that in jejunum and ileum. The jejunal and ileal crypt regions showed lower specific activity than the villus cells. About 70 per cent of total reductase activity was found in cells from the upper and the mid villus fraction in each intestinal segment.
Iglesias
J
J
Departamento de Bioquímica y Biología Molecular, Universidad de Granada, Spain.
Gonzalez-Pacanowska
D
D
Caamaño
G
G
Garcia-Peregrin
E
E
D000818:Animals; D002645:Chickens; D004386:Duodenum / Q000166:cytology / Q000201:enzymology; D006903:Hydroxymethylglutaryl CoA Reductases / Q000378:metabolism*; D007082:Ileum / Q000166:cytology / Q000201:enzymology; D066298:In Vitro Techniques; D007413:Intestinal Mucosa / Q000166:cytology / Q000201:enzymology*; D007583:Jejunum / Q000166:cytology / Q000201:enzymology; D008297:Male; D009251:NADPH-Ferrihemoprotein Reductase / Q000378:metabolism; D013347:Subcellular Fractions / Q000201:enzymology
D006903:Hydroxymethylglutaryl CoA Reductases; D009251:NADPH-Ferrihemoprotein Reductase