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. 1971 Jul 1;50(1):102–120. doi: 10.1083/jcb.50.1.102

FINE STRUCTURAL LOCALIZATION OF ACYLTRANSFERASES

The Monoglyceride and α-Glycerophosphate Pathways in Intestinal Absorptive Cells

Joan A Higgins 1, Russell J Barrnett 1
PMCID: PMC2108434  PMID: 5563442

Abstract

A study of the fine structural localization of the acyltransferases of the monoglyceride and α-glycerophosphate pathways for triglyceride synthesis in the intestinal absorptive cell is reported. Glutaraldehyde-fixed tissue was found to synthesize diglyceride and triglyceride from monopalmitin and palmityl CoA, and parallel morphological studies showed the appearance of lipid droplets in the smooth endoplasmic reticulum of the absorptive cell. Glutaraldehyde-fixed tissue also synthesized triglyceride from α-glycerophosphate, although this enzyme system was more susceptible to fixation than the monoglyceride pathway acyltransferases. Cytochemical methods for the localization of free CoA were based (a) on the formation of the insoluble lanthanium mercaptide of CoA and (b) on the reduction of ferricyanide by CoA to yield ferrocyanide which forms an insoluble precipitate with manganous ions. By these methods the monoglyceride pathway acyltransferases were found to be located mainly on the inner surface of the smooth endoplasmic reticulum. The α-glycerophosphate pathway acyltransferases were localized mainly on the rough endoplasmic reticulum. Activity limited to the outer cisternae of the Golgi membranes occurred with both pathways. The possible organization of triglyceride absorption and chylomicron synthesis is discussed in view of these results.

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Selected References

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  1. BRAGDON J. H. On the composition of chyle chylomicrons. J Lab Clin Med. 1958 Oct;52(4):564–570. [PubMed] [Google Scholar]
  2. BROWN J. L., JOHNSTON J. M. THE MECHANISM OF INTESTINAL UTILIZATION OF MONOGLYCERIDES. Biochim Biophys Acta. 1964 Jun 15;84:264–274. doi: 10.1016/0926-6542(64)90055-1. [DOI] [PubMed] [Google Scholar]
  3. Bainton D. F., Farquhar M. G. Origin of granules in polymorphonuclear leukocytes. Two types derived from opposite faces of the Golgi complex in developing granulocytes. J Cell Biol. 1966 Feb;28(2):277–301. doi: 10.1083/jcb.28.2.277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brindley D. N., Hübscher G. The intracellular distribution of the enzymes catalysing the biosynthesis of glycerides in the intestinal mucosa. Biochim Biophys Acta. 1965 Dec 2;106(3):495–509. doi: 10.1016/0005-2760(65)90066-4. [DOI] [PubMed] [Google Scholar]
  5. CLARK B., HUBSCHER G. Biosynthesis of glycerides in subcellular fractions of intestinal mucosa. Biochim Biophys Acta. 1961 Jan 29;46:479–494. doi: 10.1016/0006-3002(61)90579-0. [DOI] [PubMed] [Google Scholar]
  6. CLARK B., HUBSCHIER G. Monoglyceride transa ylase of ratintestinal mucosa. Biochim Biophys Acta. 1963 Feb 19;70:43–52. doi: 10.1016/0006-3002(63)90717-0. [DOI] [PubMed] [Google Scholar]
  7. Cardell R. R., Jr, Badenhausen S., Porter K. R. Intestinal triglyceride absorption in the rat. An electron microscopical study. J Cell Biol. 1967 Jul;34(1):123–155. doi: 10.1083/jcb.34.1.123. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DAWSON A. M., ISSELBACHER K. J. Studies on lipid metabolism in the small intestine with observations on the role of bile salts. J Clin Invest. 1960 May;39:730–740. doi: 10.1172/JCI104090. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DAWSON A. M., ISSELBACHER K. J. The esterification of palmitate-1-C14 by homogenates of intestinal mucosa. J Clin Invest. 1960 Jan;39:150–160. doi: 10.1172/JCI104014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dallner G., Siekevitz P., Palade G. E. Biogenesis of endoplasmic reticulum membranes. I. Structural and chemical differentiation in developing rat hepatocyte. J Cell Biol. 1966 Jul;30(1):73–96. doi: 10.1083/jcb.30.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. ELLMAN G. L. Tissue sulfhydryl groups. Arch Biochem Biophys. 1959 May;82(1):70–77. doi: 10.1016/0003-9861(59)90090-6. [DOI] [PubMed] [Google Scholar]
  12. FOLCH J., LEES M., SLOANE STANLEY G. H. A simple method for the isolation and purification of total lipides from animal tissues. J Biol Chem. 1957 May;226(1):497–509. [PubMed] [Google Scholar]
  13. HOFMANN A. F. Thin-layer adsorption chromatography on microscope slides. Anal Biochem. 1962 Feb;3:145–149. doi: 10.1016/0003-2697(62)90104-5. [DOI] [PubMed] [Google Scholar]
  14. JOHNSTON J. M. An in vitro study of fatty acid absorption. Proc Soc Exp Biol Med. 1958 Aug-Sep;98(4):836–838. doi: 10.3181/00379727-98-24201. [DOI] [PubMed] [Google Scholar]
  15. JOHNSTON J. M., BEARDEN J. H. Phosphatidic acids as intermediates in fatty acid absorption. Arch Biochem Biophys. 1960 Sep;90:57–62. doi: 10.1016/0003-9861(60)90611-1. [DOI] [PubMed] [Google Scholar]
  16. JOHNSTON J. M., BROWN J. L. The intestinal utilization of doubly labeled alpha-monopalmitin. Biochim Biophys Acta. 1962 May 21;59:500–501. doi: 10.1016/0006-3002(62)90210-x. [DOI] [PubMed] [Google Scholar]
  17. JOHNSTON J. M. The absorption of fatty acids by the isolated intestine. J Biol Chem. 1959 May;234(5):1065–1067. [PubMed] [Google Scholar]
  18. Johnston J. M., Rao G. A., Lowe P. A., Schwarz B. E. The nature of the stimulatory role of the supernatant fraction on triglyceride synthesis by the alpha-Glycerophosphate pathway. Lipids. 1967 Jan;2(1):14–20. doi: 10.1007/BF02531994. [DOI] [PubMed] [Google Scholar]
  19. Johnston J. M., Rao G. A., Lowe P. A. The separation of the alpha-glycerophosphate and monoglyceride pathways in the intestinal biosynthesis of triglycerides. Biochim Biophys Acta. 1967 Jun 6;137(3):578–580. doi: 10.1016/0005-2760(67)90140-3. [DOI] [PubMed] [Google Scholar]
  20. KAY D., ROBINSON D. S. The structure of chylomicra obtained from the thoracic duct of the rat. Q J Exp Physiol Cogn Med Sci. 1962 Jul;47:258–261. doi: 10.1113/expphysiol.1962.sp001605. [DOI] [PubMed] [Google Scholar]
  21. Kern F., Jr, Borgström B. Quantitative study of the pathways of triglyceride synthesis by hamster intestinal mucosa. Biochim Biophys Acta. 1965 Jun 1;98(3):520–531. doi: 10.1016/0005-2760(65)90148-7. [DOI] [PubMed] [Google Scholar]
  22. Moretz R. C., Akers C. K., Parsons D. F. Use of small angle x-ray diffraction to investigate disordering of membranes during preparation for electron microscopy. I. Osmium tetroxide and potassium permanganate. Biochim Biophys Acta. 1969 Oct 14;193(1):1–11. doi: 10.1016/0005-2736(69)90052-2. [DOI] [PubMed] [Google Scholar]
  23. PALAY S. L., KARLIN L. J. An electron microscopic study of the intestinal villus. II. The pathway of fat absorption. J Biophys Biochem Cytol. 1959 May 25;5(3):373–384. doi: 10.1083/jcb.5.3.373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rao G. A., Johnston J. M. Purification and properties of triglyceride synthetase from the intestinal mucosa. Biochim Biophys Acta. 1966 Dec 7;125(3):465–473. doi: 10.1016/0005-2760(66)90035-x. [DOI] [PubMed] [Google Scholar]
  25. Rao G. A., Johnston J. M. Studies of the formation and utilization of bound CoA in glyceride biosynthesis. Biochim Biophys Acta. 1967 Aug 8;144(1):25–33. doi: 10.1016/0005-2760(67)90074-4. [DOI] [PubMed] [Google Scholar]
  26. Rodgers J. B., Jr Assay of acyl-CoA:monoglyceride acyltransferase from rat small intestine using continuous recording spectrophotometry. J Lipid Res. 1969 Jul;10(4):427–432. [PubMed] [Google Scholar]
  27. SENIOR J. R. INTESTINAL ABSORPTION OF FATS. J Lipid Res. 1964 Oct;5:495–521. [PubMed] [Google Scholar]
  28. SENIOR J. R., ISSELBACHER K. J. Activation of long-chain fatty acids by rat-gut mucosa. Biochim Biophys Acta. 1960 Nov 4;44:399–400. doi: 10.1016/0006-3002(60)91594-8. [DOI] [PubMed] [Google Scholar]
  29. SENIOR J. R., ISSELBACHER K. J. Direct esterification of monoglycerides with palmityl coenzyme A by intestinal epithelial subcellular fractions. J Biol Chem. 1962 May;237:1454–1459. [PubMed] [Google Scholar]
  30. SRERE P. A., SEUBERT W., LYNEN F. Palmityl coenzyme A deacylase. Biochim Biophys Acta. 1959 Jun;33(2):313–319. doi: 10.1016/0006-3002(59)90118-0. [DOI] [PubMed] [Google Scholar]
  31. Salpeter M. M., Zilversmit D. B. The surface coat of chylomicrons: electron microscopy. J Lipid Res. 1968 Mar;9(2):187–192. [PubMed] [Google Scholar]
  32. Strauss E. W. Electron microscopic study of intestinal fat absorption in vitro from mixed micelles containing linolenic acid, monoolein, and bile salt. J Lipid Res. 1966 Mar;7(2):307–323. [PubMed] [Google Scholar]

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