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Proteolytic cleavage of protein kinase Cμ upon induction of apoptosis in U937 cells: Identification of the cleavage site and characterization of the fragment

Treatment of U937 cells with various apoptosis-inducing agents, such as TNFα and β-D-arabinofuranosylcytosine (ara-C) alone or in combination with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), bryostatin 1 or cycloheximide, causes proteolytic cleavage of protein kinase Cμ (PKCμ) betw...

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Bibliographic Details
Main Authors: Häussermann, Sabine (Author) , Kittstein, Walter (Author) , Rincke, Gabriele (Author) , Johannes, Franz-Josef (Author) , Marks, Friedrich (Author) , Gschwendt, Michael (Author)
Format: Article (Journal)
Language:English
Published: 30 November 1999
In: FEBS letters
Year: 1999, Volume: 462, Issue: 3, Pages: 442-446
ISSN:1873-3468
DOI:10.1016/S0014-5793(99)01577-X
Online Access:Verlag, lizenzpflichtig, Volltext: https://doi.org/10.1016/S0014-5793(99)01577-X
Verlag, lizenzpflichtig, Volltext: https://www.sciencedirect.com/science/article/pii/S001457939901577X
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Author Notes:Sabine Häussermann, Walter Kittstein, Gabriele Rincke, Franz-Josef Johannes, Friedrich Marks, Michael Gschwendt
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Summary:Treatment of U937 cells with various apoptosis-inducing agents, such as TNFα and β-D-arabinofuranosylcytosine (ara-C) alone or in combination with the phorbol ester 12-O-tetradecanoylphorbol-13-acetate (TPA), bryostatin 1 or cycloheximide, causes proteolytic cleavage of protein kinase Cμ (PKCμ) between the regulatory and catalytic domain, generating a 62 kDa catalytic fragment of the kinase. The formation of this fragment is effectively suppressed by the caspase-3 inhibitor Z-DEVD-FMK. In accordance with these in vivo data, treatment of recombinant PKCμ with caspase-3 in vitro results also in the generation of a 62 kDa fragment (p62). Treatment of several aspartic acid to alanine mutants of PKCμ with caspase-3 resulted in an unexpected finding. PKCμ is not cleaved at one of the typical cleavage sites containing the motif DXXD but at the atypical site CQND378/S379. The respective fragment (amino acids 379-912) was expressed in bacteria as a GST fusion protein (GST-p62) and partially purified. In contrast to the intact kinase, the fragment does not respond to the activating cofactors TPA and phosphatidylserine and is thus unable to phosphorylate substrates effectively.
Item Description:Gesehen am 22.04.2021
Physical Description:Online Resource
ISSN:1873-3468
DOI:10.1016/S0014-5793(99)01577-X

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