Molecular Modeling and Site-Directed Mutagenesis Studies of a Phorbol Ester-Binding Site in Protein Kinase C

1996 ◽  
Vol 39 (13) ◽  
pp. 2541-2553 ◽  
Author(s):  
Shaomeng Wang ◽  
Marcelo G. Kazanietz ◽  
Peter M. Blumberg ◽  
Victor E. Marquez ◽  
G. W. A. Milne
Keyword(s):  
Protein Kinase C ◽  
Molecular Modeling ◽  
Protein Kinase ◽  
Binding Site ◽  
Phorbol Ester ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Kinase C

scholarly journals Residues in the Second Cysteine-rich Region of Protein Kinase C δ Relevant to Phorbol Ester Binding as Revealed by Site-directed Mutagenesis

1995 ◽  
Vol 270 (37) ◽  
pp. 21852-21859 ◽  
Author(s):  
Marcelo G. Kazanietz ◽  
Shaomeng Wang ◽  
George W. A. Milne ◽  
Nancy E. Lewin ◽  
Howard L. Liu ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Phorbol Ester ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Rich Region ◽  
Kinase C

Probing the Binding of Indolactam-V to Protein Kinase C through Site-Directed Mutagenesis and Computational Docking Simulations

1999 ◽  
Vol 42 (18) ◽  
pp. 3436-3446 ◽  
Author(s):  
Shaomeng Wang ◽  
Ming Liu ◽  
Nancy E. Lewin ◽  
Patricia S. Lorenzo ◽  
Dipak Bhattacharrya ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Computational Docking ◽  
Docking Simulations ◽  
Kinase C

scholarly journals Analysis of Gz alpha by site-directed mutagenesis. Sites and specificity of protein kinase C-dependent phosphorylation.

1993 ◽  
Vol 268 (5) ◽  
pp. 3494-3498
Author(s):  
K.M. Lounsbury ◽  
B. Schlegel ◽  
M. Poncz ◽  
L.F. Brass ◽  
D.R. Manning
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Kinase C

scholarly journals Annexin V inhibits protein kinase C activity via a mechanism of phospholipid sequestration

1998 ◽  
Vol 330 (3) ◽  
pp. 1277-1282 ◽  
Author(s):  
Thierry DUBOIS ◽  
Jean-Paul MIRA ◽  
Denis FELIERS ◽  
Egle SOLITO ◽  
Françoise RUSSO-MARIE ◽  
...  
Keyword(s):  
Protein Kinase C ◽  
Protein Kinase ◽  
Binding Site ◽  
Annexin V ◽  
Jurkat Cells ◽  
Site Directed Mutagenesis ◽  
Kinase C ◽  
Protein Kinase C Activity ◽  
Mechanism Of Inhibition

In this study, we assessed the role of annexin V, a Ca2+-dependent phospholipid-binding protein, as a regulator of protein kinase C (PKC) and characterized its mechanism of inhibition. Several mutants obtained by oligonucleotide site-directed mutagenesis were tested in vitro on PKC activity in cytosolic fractions from Jurkat cells and on purified PKCα. Annexin V inhibited phosphorylation of annexin II by endogenous PKC and phosphorylation of myelin basic protein by PKCα. In both systems, the use of single Ca2+-binding-site mutants of annexin V led to a partial reversal of inhibition, and the Ca2+-binding site located in the first domain of annexin V was found to have the most important role. An increase in the number of mutated Ca2+-binding sites led to a greater loss of inhibition. These results corroborated those showing the progressive loss of binding of these mutants to phospholipid liposomes. In conclusion, we show that PKC inhibition by annexin V is the consequence of a mechanism involving phospholipid sequestration by annexin V, and that the Ca2+-binding site located in domain 1 of annexin V plays a predominant role in this process. In addition, we show that the R122AIK site, which may act analogously to a PKC-inhibitory pseudosubstrate site, is not involved in PKC inhibition, and that a peptide corresponding to the C-terminal tail of annexin V inhibits PKC activity but to a lesser extent than annexin V itself.

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