The ATP-depleting reagent "iodacetamide" induces the degradation of protein kinase C alpha (PKCα) in LLC-PK1 pig kidney cells

Adnan Dibas, Abdul J. Mia, Thomas Yorio

Research output: Contribution to journalArticle

Abstract

The alkylating reagent iodoacetamide, a potent inhibitor of sulfhydryl proteases, was found to stimulate the selective degradation of protein kinase C alpha (PKCα) isoform. Treatment of LLC-PK1 cells with iodoacetamide (5 mM) for 30-90 minutes at room temperature followed by western blotting on total cell homogenate, revealed the absence of an 80 KDa protein and the appearance of an 47 KDa band that was still recognized with the antibody. Serine protease inhibitor, metalloprotease inhibitors and leupeptin failed to prevent the degradation of PKCα. The degradation persisted at 4 °C and in the absence of Ca2+. Iodoacetamide had no direct effect on purified PKCα. In conclusion, the degradation of PKCα is a novel phenomenon. The degradation process could not be inhibited by known protease inhibitors or in the absence of Ca2+ or at 4 °C and appears to involve interactions with an unknown intermediates.

Original languageEnglish
JournalFASEB Journal
Volume11
Issue number3
StatePublished - 1 Dec 1997

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Protein Kinase C-alpha
protein kinase C
kidney cells
Iodoacetamide
Swine
Adenosine Triphosphate
Kidney
Degradation
swine
degradation
Protease Inhibitors
proteinase inhibitors
LLC-PK1 Cells
Serine Proteinase Inhibitors
Metalloproteases
calcium
metalloproteinases
Corrosion inhibitors
serine proteinases
Protein Isoforms

Cite this

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abstract = "The alkylating reagent iodoacetamide, a potent inhibitor of sulfhydryl proteases, was found to stimulate the selective degradation of protein kinase C alpha (PKCα) isoform. Treatment of LLC-PK1 cells with iodoacetamide (5 mM) for 30-90 minutes at room temperature followed by western blotting on total cell homogenate, revealed the absence of an 80 KDa protein and the appearance of an 47 KDa band that was still recognized with the antibody. Serine protease inhibitor, metalloprotease inhibitors and leupeptin failed to prevent the degradation of PKCα. The degradation persisted at 4 °C and in the absence of Ca2+. Iodoacetamide had no direct effect on purified PKCα. In conclusion, the degradation of PKCα is a novel phenomenon. The degradation process could not be inhibited by known protease inhibitors or in the absence of Ca2+ or at 4 °C and appears to involve interactions with an unknown intermediates.",
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The ATP-depleting reagent "iodacetamide" induces the degradation of protein kinase C alpha (PKCα) in LLC-PK1 pig kidney cells. / Dibas, Adnan; Mia, Abdul J.; Yorio, Thomas.

In: FASEB Journal, Vol. 11, No. 3, 01.12.1997.

Research output: Contribution to journalArticle

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T1 - The ATP-depleting reagent "iodacetamide" induces the degradation of protein kinase C alpha (PKCα) in LLC-PK1 pig kidney cells

AU - Dibas, Adnan

AU - Mia, Abdul J.

AU - Yorio, Thomas

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