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Titolo:
DEVELOPMENT AND APPLICATION OF A FLUOROMETRIC ASSAY FOR MAMMALIAN MEMBRANE DIPEPTIDASE
Autore:
HEYWOOD SP; HOOPER NM;
Indirizzi:
UNIV LEEDS,DEPT BIOCHEM & MOLEC BIOL LEEDS LS2 9JT W YORKSHIRE ENGLAND UNIV LEEDS,DEPT BIOCHEM & MOLEC BIOL LEEDS LS2 9JT W YORKSHIRE ENGLAND
Titolo Testata:
Analytical biochemistry
fascicolo: 1, volume: 226, anno: 1995,
pagine: 10 - 14
SICI:
0003-2697(1995)226:1<10:DAAOAF>2.0.ZU;2-1
Fonte:
ISI
Lingua:
ENG
Soggetto:
RENAL DIPEPTIDASE; PHOSPHOLIPASE-C; THIENAMYCIN; ENZYMES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
13
Recensione:
Indirizzi per estratti:
Citazione:
S.P. Heywood e N.M. Hooper, "DEVELOPMENT AND APPLICATION OF A FLUOROMETRIC ASSAY FOR MAMMALIAN MEMBRANE DIPEPTIDASE", Analytical biochemistry, 226(1), 1995, pp. 10-14

Abstract

Membrane dipeptidase (EC 3.4.13.19) is a widely distributed mammaliancell surface enzyme involved in the metabolism of glutathione, leukotriene D-4, and certain beta-lactam antibiotics. In this study we have developed a sensitive and rapid assay for membrane dipeptidase based on the fluorometric detection of the D-Phe released from the model substrate Gly-D-Phe. The released D-Phe is first acted on by D-amino acid oxidase in the presence of flavin adenine dinucleotide. The resulting hydrogen peroxide is then metabolized by peroxidase in the presence ofthe acceptor substrate p-hydroxyphenylacetic acid which is converted to a highly fluorescent compound. The assay configuration is sensitivedown to 0.1 nmol D-Phe and can accurately measure membrane dipeptidase activity even in the presence of large amounts of contaminating protein. The membrane dipeptidase assay and the subsequent fluorometric detection of the released D-Phe can be performed in microtiter plates, thus taking less than 1 h to process 96 samples. This sensitive and rapid assay will be useful for the routine measurement of membrane dipeptidase activity in a number of different applications. (C) 1995 Academic Press, Inc.

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Documento generato il 03/12/20 alle ore 21:52:09