Abstract
Incorporation of L-[U-14C]leucine and of D[U-14C]glucose into proteins of fresh human gastric mucosa in vitro was studied after incubation of homogenized tissue and of intact mucosal pieces. CsCl centrifugation was used to separate high-density mucus glycoproteins from other mucosal proteins, and the macromolecular nature of radioactive mucosal glycoprotein fractions was confirmed by SDS/polyacrylamide-gel electrophoresis and autoradiography of the polyacrylamide gels. In all experiments a substantial proportion of total incorporated radioactivity was associated with gastric-mucosal glycoprotein fractions (CsCl fraction L3), indicating their biosynthesis. Radioactivity of these fractions was shown to co-chromatograph with carbohydrates when fractionated either directly or after reduction and alkylation (1) Sephadex G-200 chromatography in the excluded fractions and (2) by DEAE-cellulose ion-exchange chromatography. On incubation of intact mucosa, the major portion of radioactivity associated with the glycoprotein fractions of both leucine- and glucose-labelled specimens was secreted into the mucosal media during the course of the experiment. It is suggested that biosynthesis of mucus in vivo by gastric mucosa may be associated with rapid secretion of the synthesized macromolecules into the lumen of the stomach and that investigations of the metabolic processes within the mucosa should consider the products of secretion of the tissue. Incorporation of L-[U-14C]leucine implies biosynthesis of the polypeptide components of the macromolecules.
Full text
PDF







Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bhaskar K. R., Creeth J. M. The macromolecular properties of blood-group-specific glycoproteins. Characterization of a series of fractions obtained by density-gradient ultracentrifugation. Biochem J. 1974 Dec;143(3):669–679. doi: 10.1042/bj1430669. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carlstedt I., Sheehan J. K., Corfield A. P., Gallagher J. T. Mucous glycoproteins: a gel of a problem. Essays Biochem. 1985;20:40–76. [PubMed] [Google Scholar]
- Contractor S. F., Shane B. Purification and characterization of lysosomal -glucuronidase from human placenta. Biochem J. 1972 Jun;128(1):11–18. doi: 10.1042/bj1280011. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dekanski J. B., Macdonald A., Sacra P. Effects of fasting, stress and drugs on gastric glycoprotein synthesis in the rat. Br J Pharmacol. 1975 Nov;55(3):387–392. doi: 10.1111/j.1476-5381.1975.tb06942.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Jentjens T., Smits H. L., Strous G. J. 16,16-Dimethyl prostaglandin E2 stimulates galactose and glucosamine but not serine incorporation in rat gastric mucous cells. Gastroenterology. 1984 Aug;87(2):409–416. [PubMed] [Google Scholar]
- Jentjens T., Strous G. J. Quantitative aspects of mucus glycoprotein biosynthesis in rat gastric mucosa. Biochem J. 1985 May 15;228(1):227–232. doi: 10.1042/bj2280227. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kent P. W., Allen A. The biosynthesis of intestinal mucins. The effect of salicylate on glycoprotein biosynthesis by sheep colonic and human gastric mucosal tissues in vitro. Biochem J. 1968 Feb;106(3):645–658. doi: 10.1042/bj1060645. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lukie B. E., Forstner G. G. Synthesis of intestinal clycoproteins. Inhibition of (I- 14 C)glucosamine incorporation by sodium salicylate in vitro. Biochim Biophys Acta. 1972 Jul 19;273(2):380–388. doi: 10.1016/0304-4165(72)90230-9. [DOI] [PubMed] [Google Scholar]
- Parke D. V., Symons A. M. The biochemical pharmacology of mucus. Adv Exp Med Biol. 1977;89:423–441. doi: 10.1007/978-1-4613-4172-7_29. [DOI] [PubMed] [Google Scholar]
- Porter P. Studies of porcine secretory IgA and its component chains in relation to intestinal absorption of colostral immunoglobulins by the neonatal pig. Immunology. 1973 Jan;24(1):163–176. [PMC free article] [PubMed] [Google Scholar]
- Slomiany A., Slomiany B. L., Witas H., Aono M., Newman L. J. Isolation of fatty acids covalently bound to the gastric mucus glycoprotein of normal and cystic fibrosis patients. Biochem Biophys Res Commun. 1983 May 31;113(1):286–293. doi: 10.1016/0006-291x(83)90464-3. [DOI] [PubMed] [Google Scholar]
- Snary D., Allen A. Studies on gastric mucoproteins. The isolation and characterization of the mucoprotein of the water-soluble mucus from pig cardiac gastric mucosa. Biochem J. 1971 Aug;123(5):845–853. doi: 10.1042/bj1230845. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Snary D., Allen A. Studies on gastric mucoproteins. The production of radioactive mucoproteins by pig gastric mucosal scrapings in vitro. Biochem J. 1972 Apr;127(3):577–587. doi: 10.1042/bj1270577. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spohn M., McColl I. Ethanol, its effect on the synthesis of proteins by guinea-pig gastric mucosa. Biochem Pharmacol. 1986 Jun 1;35(11):1909–1914. doi: 10.1016/0006-2952(86)90311-4. [DOI] [PubMed] [Google Scholar]
- Spohn M., McColl I. In vitro studies on the effect of salicylates on the synthesis of proteins by guinea pig gastric mucosal tissue. Biochim Biophys Acta. 1980 Jul 29;608(2):409–421. doi: 10.1016/0005-2787(80)90186-0. [DOI] [PubMed] [Google Scholar]
- Spohn M., McColl I. Studies on gastric mucosal IgA: separation of immunoglobulin rich fraction from gastric mucoproteins. Biochem Biophys Res Commun. 1977 Dec 7;79(3):837–842. doi: 10.1016/0006-291x(77)91187-1. [DOI] [PubMed] [Google Scholar]
- Starkey B. J., Snary D., Allen A. Characterization of gastric mucoproteins isolated by equilibrium density-gradient centrifugation in caesium chloride. Biochem J. 1974 Sep;141(3):633–639. doi: 10.1042/bj1410633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Trotman C. N., Greenwell J. R. Computed resolution and relative specific radioactivities of radiolabelled proteins synthesized by isolated gastric mucosal cells. Biochem J. 1979 Jan 15;178(1):159–164. doi: 10.1042/bj1780159. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Witas H., Sarosiek J., Aono M., Murty V. L., Slomiany A., Slomiany B. L. Lipids associated with rat small-intestinal mucus glycoprotein. Carbohydr Res. 1983 Aug 16;120:67–76. doi: 10.1016/0008-6215(83)88007-0. [DOI] [PubMed] [Google Scholar]
- Yeomans N. D., Millar S. J. Synthesis and secretion of glycoproteins by mucosa of rat gastric antrum in organ culture. Dig Dis Sci. 1980 Apr;25(4):295–301. doi: 10.1007/BF01308521. [DOI] [PubMed] [Google Scholar]
- Zacharius R. M., Zell T. E., Morrison J. H., Woodlock J. J. Glycoprotein staining following electrophoresis on acrylamide gels. Anal Biochem. 1969 Jul;30(1):148–152. doi: 10.1016/0003-2697(69)90383-2. [DOI] [PubMed] [Google Scholar]

