Abstract
The destruction of 125I-labelled insulin by an enzyme system from rat adipose tissue was studied. The system was located in the particulate fraction. Activity was assayed by the amount of 125I-labelled degradation products rendered soluble in trichloroacetic acid. The system was heat-labile, with an alkaline pH optimum. The velocity of the reaction varied directly with the enzyme concentration. Paper chromatography of the degradation products showed six ninhydrin-sensitive areas with radioactivity coinciding with three of them. Albumin inhibited the system; ribonuclease did not. Although only 25% of the total 125I-label was detected by this assay, results with insulin-specific assays suggested that most (80–90%) of the hormone was inactivated. Possible interpretations of these results are discussed. The particulate fractions of other tissues were also studied.
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- BALL E. G., MERRILL M. A. A manometric assay of insulin and some results of the application of the method to sera and islet-containing tissues. Endocrinology. 1961 Sep;69:596–607. doi: 10.1210/endo-69-3-596. [DOI] [PubMed] [Google Scholar]
- Carpenter F. H. Relationship of structure to biological activity of insulin as revealed by degradative studies. Am J Med. 1966 May;40(5):750–758. doi: 10.1016/0002-9343(66)90156-2. [DOI] [PubMed] [Google Scholar]
- Crofford O. B. The uptake and inactivation of native insulin by isolated fat cells. J Biol Chem. 1968 Jan 25;243(2):362–369. [PubMed] [Google Scholar]
- De Duve C., Wattiaux R. Functions of lysosomes. Annu Rev Physiol. 1966;28:435–492. doi: 10.1146/annurev.ph.28.030166.002251. [DOI] [PubMed] [Google Scholar]
- Garratt C. J., Jarrett R. J., Keen H. The relationship between insulin association with tissues and insulin action. Biochim Biophys Acta. 1966 May 26;121(1):143–150. doi: 10.1016/0304-4165(66)90357-6. [DOI] [PubMed] [Google Scholar]
- HALES C. N., RANDLE P. J. Immunoassay of insulin with insulin-antibody precipitate. Biochem J. 1963 Jul;88:137–146. doi: 10.1042/bj0880137. [DOI] [PMC free article] [PubMed] [Google Scholar]
- NARAHARA H. T., TOMIZAWA H. H., MILLER R., WILLIAMS R. H. Intracellular distribution of an insulin-inactivating system of liver. J Biol Chem. 1955 Dec;217(2):675–684. [PubMed] [Google Scholar]
- SPRINGELL P. H. An unreactive tyrosine residue in insulin and the exclusive iodination of the A chain. Nature. 1961 Sep 30;191:1372–1373. doi: 10.1038/1911372a0. [DOI] [PubMed] [Google Scholar]
- TOMIZAWA H. H. Mode of action of an insulin-degrading enzyme from beef liver. J Biol Chem. 1962 Feb;237:428–431. [PubMed] [Google Scholar]
- TOMIZAWA H. H. Properties of glutathione insulin transhydrogenase from beef liver. J Biol Chem. 1962 Nov;237:3393–3396. [PubMed] [Google Scholar]
- Westman S. Degradation of insulin in vitro by liver and epididymal adipose tissue from obese-hyperglycaemic mice. Biochem J. 1968 Jan;106(2):543–547. doi: 10.1042/bj1060543. [DOI] [PMC free article] [PubMed] [Google Scholar]