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Localisation of proline oxidase and Δ-pyrroline-5-carboxylic acid dehydrogenase in rat liver

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Keywords
  • Medizin
  • Ddc:610
Disciplines
  • Biology
  • Communication

Abstract

Localisation of proline oxidase and Δ-pyrroline-5-carboxylic acid dehydrogenase in rat liver Volume 3, number 4 FEBS LETTERS June 1969 LOCALISATION OF PROLINE OXIDASE AND A-PYRROLINE-S-CARBOXYLIC ACID DEHYDROGENASE IN RAT LIVER G.BRUNNER and W.NEUPERT InstirU t fiir Physiologische Chemie und Physikalische Biochemie der Universitdt M&hen Received 26 April 1969 1. Introduction The enzymes of prolinsglutamate oxidation and the preparation of their substrates have been de- scribed by Strecker and coworkers. [ 1,3] . The pres- ence of proline oxidase in rat liver mitochondria was described by Johnson and Strecker [2]. A rough lo- calisation of proline oxidase and A-pyrroline&carbox- ylic acid dehydrogenase in locust flight muscle has been attempted by Brosemer and Veerabhadrappa [4]. Lang and Lang [lo] reported proline oxidase to be bound to insoluble particles in rat liver cells. This communication describes the localisation of both enzymes in rat liver by the methods of frac- tionated extraction [ 5,6] and an improved method of preparing inner and outer membranes of rat liver mitochondria. 2. Materials and methods 2.1. Fmctionated extraction Rat liver tissue was fractionated by the method of Delbrtick et al. [S] and Pette [6] which yields primarily a separation of extra- and intramitochon- drial enzymes. Abbreviations used: FCDH = A-pyrroline-5-carboxylic acid dehydrogenase MDH = malate dehydrogenase SDH = succinic dehydrogenase GLUDH = glutamate dehydrogenase MAO = monoamine oxidase. ADK = adenylate kinase TRA = triethanolamine liydrochloride 2.2. Preparation of highly purified inner and outer mitochondrial mitochondria were prepared and treated as previously described by Neupert et al. [7], with the following modifications. After the step of swelling and shrinking, the mitochondrial discontinuous sucrose gradient of total volume 50 ml, increasing from 30% to 55% by 2.5% at each step. After centrifugation using the Be

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