Summary
A low molecular weight fraction of human urine (<500 daltons) which both inhibits monoamine oxidase and benzodiazepine binding to central and peripheral receptors has been purified by ethyl acetate extractions, HPLC and thin layer chromatography. This material extracted equally well at acid and basic pH and was insoluble in heptane. It competitively inhibited binding of3H-clonazepam, a central benzodiazepine receptor agonist and, in addition, displaced3H-Ro 5-4864, a specific peripheral benzodiazepine receptor ligand, from its binding sites. It showed no GABA shift with the benzodiazepine receptor antagonist, Ro-15 1788. MAO A and B were inhibited approximately equipotently and the material competitively inhibited tyramine oxidation by rat liver. It was stable on boiling and is unlikely to be a peptide.
Similar content being viewed by others
References
Armando I, Glover V, Sandler M (1986) Distribution of endogenous benzodiazepine receptor ligand-monoamine oxidase inhibitory activity (tribulin) in tissues. Life Sci 38: 2063–2067
Beaumont K, Cheung AK, Geller ML, Fanestil DD (1983) Inhibitors of periipheral-type benzodiazepine receptors present in human urine and plasma ultrafiltrates. Life Sci 33: 1375–1384
Becker RE, Giambalvo C, Fox RA, Macho M (1983) Endogenous inhibitors of monoamine oxidase present in human cerebrospinal fluid. Science 221: 476–478
Bhattacharya SK, Glover V, Sandler M, Clow A, Topham A, Bernadt M, Murray R (1982) Raised endogenous monoamine oxidase inhibitor output in postwithdrawal alcoholics: effects of L-dopa and ethanol. Biol Psychiat 17: 829–836
Braestrup C, Nielsen M, Olsen CE (1980) Urinary and brainβ-carboline-3carboxylates as potent inhibitors of brain benzodiazepine receptors. Proc Natl Acad Sci, U.S.A. 77: 2288–2292
Clow A, Glover V, Armando I, Sandler M (1983) New endogenous benzodiazepine receptor ligand in human urine: identity with endogenous monoamine oxidase inhibitor? Life Sci 33: 735–741
Davis LG, Cohen RK (1980) Identification of an endogenous peptide-ligand for the benzodiazepine receptor. Biochem Biophys Res Commun 92: 141–148
Dorow R, Horowski R, Paschelke G, Amin M, Braestrup C (1983) Severe anxiety induced by FG 7142, aβ-carboline ligand for benzodiazepine receptors. Lancet ii: 98–99
Fehske KJ, Muller WE, Platt KL, Stillbauer A (1981) Inhibition of benzodiazepine receptor binding by several tryptophan and indole derivatives. Biochem Pharmacol 30: 3016–3019
Giambalvo CT, Becker RE (1981) Modulators of monoamine oxidase in plasma. Life Sci 29: 2017–2024
Glover V, Bhattacharya SK, Sandler M, File SE (1981) Benzodiazepines reduce stress-augmented increase in rat urine monoamine oxidase inhibitor. Nature 292: 347–349
Glover V, Liebowitz J, Armando I, Sandler M (1982)β-Carbolines as selective monoamine oxidase inhibitors:in vivo implications. J Neural Transm 54: 209–218
Glover V, Reveley MA, Sandler M (1980) A monoamine oxidase inhibitor in human urine. Biochem Pharmacol 29: 467–470
Glover V, Segarajasinghe CE, Clow A, Elsworth JD, Sandler M (1984) Endogenous monoamine oxidase inhibitors in rat brain and human plasma. J Pharm Pharmacol 36: 12 W
Guidotti A, Forchetti CM, Corda MG, Konkel D, Bennett CD, Costa E (1983) Isolation, characterization, and purification to homogeneity of an endogenous polypeptide with agonistic action of benzodiazepine receptors. Proc Natl Acad Sci USA 80: 3531–3533
Karobath M, Sperk G, Schonbeck G (1978) Evidence for an endogenous factor interfering with3H-diazepam binding to rat brain membrane. Eur J Pharmacol 49: 323–326
Kuhn W, Neuser D, Przuntek H (1981) [3H]Diazepam displacing activity in human cerebrospinal fluid. J Neurochem 37: 1045–1047
Lewinsohn R, Bohm K-H, Glover V, Sandler M (1978) A benzylamine oxidase distinct from monoamine oxidase B — widespread distribution in man and rat. Biochem Pharmacol 27: 1857–1863
Mantione CR, Goldman ME, Weissman BA, Paul SM, Skolnick P (1985) Endogenous inhibitors of [3H] Ro 5-4864 binding to “peripheral-type” binding sites for benzodiazepines are present in peripheral tissues and brain. In: Lal H, LaBella F, Lane J (eds) Endocoids. Alan R Liss, New York, pp 167–173
Marangos PJ, Patel J, Skolnick P, Paul SM (1982) Endogenous “benzodiazepine-like” agents. In: Usdin E, Skolnick P, Tallman JF, Paul SM (eds) Pharmacology of benzodiazepines. Macmillan, London, pp 519–528
Petursson H, Bhattacharya SK, Glover V, Sandler M, Lader MH (1982) Urinary monoamine oxidase inhibitor and benzodiazepine withdrawal. Br J Psychiat 140: 7–10
Sandler M (1982) The emergence of tribulin. Trends Pharmac Sci 3: 471–472
Skolnick P, Schweri MM, Evans EF, Moncada VY, Paul SM (1982) Anin vitro binding assay which differentiates benzodiazepine ‘agonists’ and ‘antagonists’. Eur J Pharmacol 78: 133–136
Smith I (ed) (1958) Chromatographic techniques. Heinemann, London
Wu J-Y, Lin HS, Su YYT, Yang CY (1985) Isolation and purification of an endogenous brain ligand for benzodiazepine receptor(s). In: Lal H, LaBella F, Lane J (eds) Endocoids. Alan R Liss, New York, pp 485–493
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Elsworth, J.D., Dewar, D., Glover, V. et al. Purification and characterization of tribulin, an endogenous inhibitor of monoamine oxidase and of benzodiazepine receptor binding. J. Neural Transmission 67, 45–56 (1986). https://doi.org/10.1007/BF01243358
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01243358