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. 1974 Aug;71(8):3217–3220. doi: 10.1073/pnas.71.8.3217

Studies on the Carbohydrate Moiety of Dopamine β-Hydroxylase: Interaction of the Enzyme with Concanavalin A

Ellen F Wallace 1, Walter Lovenberg 1
PMCID: PMC388654  PMID: 4607118

Abstract

Dopamine β-hydroxylase [EC 1.14.17.1; 3,4-dihydroxyphenylethylamine, ascorbate:oxygen oxidoreductase (β-hydroxylating)], a glycoprotein, was found to interact strongly with the plant lectin, concanavalin A. This interaction did not appear to alter the catalytic properties of the enzyme and did not prevent interaction of the protein with specific antibodies. Dopamine-β-hydroxylase is adsorbed to concanavalin A covalently bound to Sepharose and can be eluted with α-methylmannoside. The enzyme is catalytically active when it is immobilized on a concanavalin A-Sepharose column. Enzymic removal of four of seven mannose residues present in the monomer does not result in the loss of any catalytic activity. It is concluded that the active site is not near the point of carbohydrate attachment on the molecule and that this may be of significance in orienting the enzyme on the vesicular membrane.

Keywords: glycoenzyme, chromaffin vesicle, norepinephrine, adrenal, lectin

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belpaire F., Laduron P. Tissue fractionation and catecholamines. 3. Intracellular distribution of endogenous inhibitors of dopamine- -hydroxylase in adrenal medulla. Biochem Pharmacol. 1970 Apr;19(4):1323–1331. doi: 10.1016/0006-2952(70)90047-x. [DOI] [PubMed] [Google Scholar]
  2. CREVELING C. R., VAN DER SCHOOT J. B., UDENFRIEND S. Phenethylamine isosteres as inhibitors of dopamine beta-oxidase. Biochem Biophys Res Commun. 1962 Jul 3;8:215–219. doi: 10.1016/0006-291x(62)90266-8. [DOI] [PubMed] [Google Scholar]
  3. Foldes A., Jeffrey P. L., Preston B. N., Austin L. Dopamine -hydroxylase of bovine adrenal medullae. A rapid purification procedure. Biochem J. 1972 Mar;126(5):1209–1217. doi: 10.1042/bj1261209. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Friedman S., Kaufman S. 3,4-dihydroxyphenylethylamine beta-hydroxylase. Physical properties, copper content, and role of copper in the catalytic acttivity. J Biol Chem. 1965 Dec;240(12):4763–4773. [PubMed] [Google Scholar]
  5. GOLDSTEIN I. J., HOLLERMAN C. E., MERRICK J. M. PROTEIN-CARBOHYDRATE INTERACTION. I. THE INTERACTION OF POLYSACCHARIDES WITH CONCANAVALIN A. Biochim Biophys Acta. 1965 Jan 4;97:68–76. doi: 10.1016/0304-4165(65)90270-9. [DOI] [PubMed] [Google Scholar]
  6. 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]
  7. Lee C. Y., Scocca J. R. A common structural unit in asparagine-oligosaccharides of several glycoproteins from different sources. J Biol Chem. 1972 Sep 25;247(18):5753–5758. [PubMed] [Google Scholar]
  8. Rush R. A., Thomas P. E., Kindler S. H., Udenfriend S. The interaction of dopamine-beta-hydroxylase with concanavalin A and its use in enzyme purification. Biochem Biophys Res Commun. 1974 Apr 23;57(4):1301–1305. doi: 10.1016/0006-291x(74)90837-7. [DOI] [PubMed] [Google Scholar]
  9. Spiro R. G. Glycoproteins. Annu Rev Biochem. 1970;39:599–638. doi: 10.1146/annurev.bi.39.070170.003123. [DOI] [PubMed] [Google Scholar]
  10. Sukeno T., Tarentino A. L., Plummer T. H., Jr, Maley F. Purification and properties of -D- and -D-mannosidases from hen oviduct. Biochemistry. 1972 Apr 11;11(8):1493–1501. doi: 10.1021/bi00758a026. [DOI] [PubMed] [Google Scholar]
  11. Tarentino A. L., Plummer T. H., Jr, Maley F. The release of intact oligosaccharides from specific glycoproteins by endo-beta-N-acetylglucosaminidase H. J Biol Chem. 1974 Feb 10;249(3):818–824. [PubMed] [Google Scholar]
  12. Wallace E. F., Krantz M. J., Lovenberg W. Dopamine-beta-hydroxylase: a tetrameric glycoprotein. Proc Natl Acad Sci U S A. 1973 Aug;70(8):2253–2255. doi: 10.1073/pnas.70.8.2253. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Winkler H., Hörtnagl H., Smith A. D. Membranes of the adrenal medulla. Behaviour of insoluble proteins of chromaffin granules on gel electrophoresis. Biochem J. 1970 Jun;118(2):303–310. doi: 10.1042/bj1180303. [DOI] [PMC free article] [PubMed] [Google Scholar]

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