The catalytic subunit of Dictyostelium cAMP-dependent protein kinase -- role of the N-terminal domain and of the C-terminal residues in catalytic activity and stability.

Details

Serval ID
serval:BIB_1405
Type
Article: article from journal or magazin.
Collection
Publications
Institution
Title
The catalytic subunit of Dictyostelium cAMP-dependent protein kinase -- role of the N-terminal domain and of the C-terminal residues in catalytic activity and stability.
Journal
European Journal of Biochemistry / Febs
Author(s)
Etchebehere L.C., Van Bemmelen M.X., Anjard C., Traincard F., Assemat K., Reymond C., Véron M.
ISSN
0014-2956 (Print)
ISSN-L
0014-2956
Publication state
Published
Issued date
1997
Volume
248
Number
3
Pages
820-826
Language
english
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: ppublish
Abstract
The C subunit of Dictyostelium cAMP-dependent protein kinase (PKA) is unusually large (73 kDa) due to the presence of 330 amino acids N-terminal to the conserved catalytic core. The sequence following the core, including a C-terminal -Phe-Xaa-Xaa-Phe-COOH motif, is highly conserved. We have characterized the catalytic activity and stability of C subunits mutated in sequences outside the catalytic core and we have analyzed their ability to interact with the R subunit and with the heat-stable protein-kinase inhibitor PKI. Mutants carrying deletions in the N-terminal domain displayed little difference in their kinetic properties and retained their capacity to be inhibited by R subunit and by PKI. In contrast, the mutation of one or both of the phenylalanine residues in the C-terminal motif resulted in a decrease of catalytic activity and stability of the proteins. Inhibition by the R subunit or by PKI were however unaffected. Sequence-comparison analysis of other protein kinases revealed that a -Phe-Xaa-Xaa-Phe- motif is present in many Ser/Thr protein kinases, although its location at the very end of the polypeptide is a particular feature of the PKA family. We propose that the presence of this motif may serve to identify isoforms of protein kinases.
Keywords
Amino Acid Sequence, Animals, Carrier Proteins/metabolism, Carrier Proteins/pharmacology, Cyclic AMP/metabolism, Cyclic AMP-Dependent Protein Kinases/chemistry, Cyclic AMP-Dependent Protein Kinases/genetics, Dictyostelium/enzymology, Electrophoresis, Polyacrylamide Gel, Enzyme Inhibitors/pharmacology, Enzyme Stability, Escherichia coli/genetics, Gene Expression, Hot Temperature, Intracellular Signaling Peptides and Proteins, Kinetics, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Denaturation, Recombinant Proteins/chemistry, Recombinant Proteins/isolation &amp, purification, Sequence Alignment, Urea
Pubmed
Web of science
Open Access
Yes
Create date
19/11/2007 12:06
Last modification date
20/08/2019 12:42
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