A New Rapid and Sensitive Bioluminescence Assay for Monoamine Oxidase Activity

Tenne, M.; Finberg, J. P. M.; Youdim, Moussa B. H.; Ulitzur, S.

doi:10.1111/j.1471-4159.1985.tb08773.x

Cited by 14 publications

(3 citation statements)

References 20 publications

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“…Following thawing of cryopreserved human thyroid cells, MAO activity was measured by the luminescence method of Tenne et al, (1985) with octylamine as the substrate. For assaying the different forms of MAO (types A and B), the radiochemical procedure described by Tipton & Youdim (1984) The assay in a total volume of 500 p1 containing 0.05M sodium phosphate buffer, pH 7.4 and 5-HT (200pM) or PEA (20pM) was incubated for 30min at 370C.…”

Section: Assay Ofmonoamine Oxidase Activitymentioning

confidence: 99%

Monoamine oxidase activity and triiodothyronine biosynthesis in human cultured thyroid cells

Kraiem

Sadeh

Youdim

1989

British J Pharmacology

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1 The proposal that monoamine oxidase (MAO) is a source of peroxide in thyroid hormone biosynthesis has been examined by use of isolated cultured human thyroid cells which retain the ability to secrete triiodothyronine (T3) in response to thyroid stimulating hormone (TSH). 2 The results demonstrated the presence of MAO A and B in human thyroid cells which oxidized 5-hydroxytryptamine and 2-phenylethylamine, respectively, and were selectively inhibited by the MAO inhibitors clorgyline and (-)-deprenyl. 3 Addition of propylthiouracil to the culture system induced a 61% reduction in TSH-stimulated T3 secretion, indicating that the bulk of such secretion apparently derives from de novo iodothyronine synthesis. 4 The MAO A and B substrate, tyramine, was ineffective in stimulating T3 secretion.

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Section: Assay Ofmonoamine Oxidase Activitymentioning

confidence: 99%

Monoamine oxidase activity and triiodothyronine biosynthesis in human cultured thyroid cells

Kraiem

Sadeh

Youdim

1989

British J Pharmacology

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“…The PRP was collected and assayed for M A 0 activity. M A 0 activity was assayed by the continuous bioluminescence method of Tenne et al (1985). Tissue homogenates (20-200 pl) were diluted to a final volume of 1 ml with sodium-potassium phosphate buffer (50 mM, pH = 7.2 at 30°C) containing 2% NaCl and 10 p1 of inhibitor solution or distilled water, as appropriate.…”

Section: A 0 Preparationsmentioning

confidence: 99%

“…These and other observations have led Fowler et al (1978) to suggest that the above binary classification may be an oversimplification. Recently, we have described the use of dark mutants of luminous bacteria for assay and monitoring of M A 0 (Tenne et al, 1985) utilising long-chain aliphatic amines as substrates. Initially we wished to characterise these amines as substrates for MAO-A or MAO-B.…”

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confidence: 99%

Deamination of Aliphatic Amines by Monoamine Oxidase A and B Studied Using a Bioluminescence Technique

Tenne

Youdim

Ulitzur

et al. 1985

Journal of Neurochemistry

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Deamination of n-octylamine and n-decylamine has been studied in various tissues using a new bioluminescence technique. Selectivity of n-octylamine and n-decylamine as substrates for monoamine oxidase (MAO) A or B has been determined using both clorgyline and (-)-deprenyl inhibition curves and kinetic parameters. Homogenates of rat brain, liver and heart containing predominantly MAO-A or -B were prepared by preincubation for 60 min with (-)-deprenyl or clorgyline (30 nM), respectively. Human placenta (MAO-A) and platelet (MAO-B) were used as reference tissues containing only one MAO form. In tissues (rat liver, brain) containing both MAO forms in equal proportion, inhibition curve studies showed a preference of both substrates for the B form of the enzyme; however, where MAO-A was the major form (rat heart, human placenta), clorgyline was the more effective inhibitor. In the beef brain cortex n-octylamine showed marked preference for MAO-B, whereas n-decylamine was selective toward-MAO-A. Kinetic studies in general supported the picture of greater selectivity of the aliphatic amine substrates for deamination by MAO-B, as reflected by lower Km values for this enzyme type. However, n-octylamine was more selective for MAO-B than n-decylamine in both kinetic and inhibition curve studies. The deamination of these aliphatic amine substrates cannot be explained only by reference to the binary classification of MAO into types A and B.

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Enzymatic Assays for High‐Throughput Screening

Carettoni

Verwaerde

2010

Burger's Medicinal Chemistry and Drug Discovery

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High‐throughput screening (HTS) is a multidisciplinary approach to investigate in parallel the pharmacological properties of a large number of small molecules, with the aim of identifying target‐specific lead compounds to be progressed as bioactive probes or therapeutic drugs. The complex integration of chemistry, biology, automation, informatics, and medicinal chemistry has prompted the evolution of HTS as an autonomous discipline integrated in the drug discovery process. Thus, HTS assays are differentiated from traditional laboratory assays by the requirement of extensive assay development and by the strict quality criteria that must be fulfilled to approach a screening campaign. In this perspective, biochemistry and enzymology have provided an essential support for the design and configuration of functional HTS assays for enzymatic targets. In turn, the impressive technological improvements of HTS have made available an unprecedented array of novel methods and instrumentations that have contributed to advance the biochemical and kinetic characterization of enzymes. In this chapter, the paradigms for the configuration of enzymatic assays for HTS are reviewed, emphasizing the tight interdependence between type and quality of the HTS assay and the outcome of a screening campaign. The sequential steps of assay development for HTS are examined, providing examples of alternative approaches to configure the activity of enzyme drug targets as HTS‐compatible assays.

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A New Rapid and Sensitive Bioluminescence Assay for Monoamine Oxidase Activity

Cited by 14 publications

References 20 publications

Monoamine oxidase activity and triiodothyronine biosynthesis in human cultured thyroid cells

Monoamine oxidase activity and triiodothyronine biosynthesis in human cultured thyroid cells

Deamination of Aliphatic Amines by Monoamine Oxidase A and B Studied Using a Bioluminescence Technique

Enzymatic Assays for High‐Throughput Screening

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