| Localization and characterization of N-ethylmaleimide sensitive inhibitor(s) of thiol cathepsin activity from cultured nil and polyoma virus-transformed nil hamster cells. | |
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MedLine Citation:
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PMID: 6267078 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Exposure of cultured Nil (a stable line of fibroblast cells from Syrian hamsters) or polyoma virus-transformed (PyNil) hamster fibroblasts to 0.5 mM N-ethylmaleimide for 5 minutes resulted in striking increases in thiol cathepsin activity in unfractionated cell-free lysates. The paradoxical increase in activity of the normally N-ethylmaleimide-sensitive cathepsins apparently occurred as the result of the protective compartmentalization of the cathepsins in the lysosomes (20,000 X g sedimented fraction) and the unprotected localization of an inhibitor(s) in the soluble cytoplasm (175,000 X g supernatant fraction). Under continuous exposure of the cells to N-ethylmaleimide, a rapid increase in cathepsin activity (seen in the first 5 minutes) was followed by a steady decrease in activity (half inactivation time, 90 minutes). The relative difference in rates of N-ethylmaleimide inactivation of thiol cathepsins and thiol cathepsin inhibitors provides a means for estimating lysosomal cathepsin activity in whole cell extracts without the need for more time-consuming fractionation procedure. In reciprocal inhibition tests, it was found that, regardless of the source of cathepsins, the Nil and PyNil cathepsin inhibitor(s) inactivated the cathepsins to approximately the same extent. The inhibitors were heat stable (90-100 degrees C for 15 minutes) at pH 4, but were totally inactivated when boiled at pH 8.5. On a calibrated Sephadex G-100 column, the relative molecular weight (Mr) of the inhibitor(s) was 13,000 daltons. On the same column, the Mr of the cathepsins was 24,000 daltons. Compared with the cathepsin activity from Nil cells, there was about five times less cathepsin activity recoverable from the PyNil cells. |
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Authors:
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R A Morgan; K L Inge; C W Christopher |
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Publication Detail:
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Type: In Vitro; Journal Article; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Journal of cellular physiology Volume: 108 ISSN: 0021-9541 ISO Abbreviation: J. Cell. Physiol. Publication Date: 1981 Jul |
Date Detail:
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Created Date: 1981-10-25 Completed Date: 1981-10-25 Revised Date: 2009-11-19 |
Medline Journal Info:
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Nlm Unique ID: 0050222 Medline TA: J Cell Physiol Country: UNITED STATES |
Other Details:
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Languages: eng Pagination: 55-66 Citation Subset: IM |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Aminopeptidases
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antagonists & inhibitors Animals Benzoylarginine-2-Naphthylamide / antagonists & inhibitors Cathepsin H Cathepsins / antagonists & inhibitors*, metabolism Cell Compartmentation Cell Fractionation Cell Line Cell Transformation, Viral Cricetinae Cysteine Endopeptidases* Cytoplasm / analysis* Ethylmaleimide / pharmacology* Hydrogen-Ion Concentration Lysosomes / enzymology* Molecular Weight Polyomavirus Temperature |
| Grant Support | |
ID/Acronym/Agency:
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GM 27905/GM/NIGMS NIH HHS; RR 07043/RR/NCRR NIH HHS |
| Chemical | |
Reg. No./Substance:
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128-53-0/Ethylmaleimide; 305-09-9/Benzoylarginine-2-Naphthylamide; EC 3.4.-/Cathepsins; EC 3.4.11.-/Aminopeptidases; EC 3.4.22.-/Cysteine Endopeptidases; EC 3.4.22.16/Cathepsin H |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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