In vitro activation of a 60–70 kDa histone H4 protein kinase from neutrophils by limited proteolysis

1996 ◽  
Vol 1295 (1) ◽  
pp. 89-95
Author(s):  
Ruichun Liu ◽  
Paul Leavis ◽  
John A. Badwey
Keyword(s):  
Protein Kinase ◽  
Limited Proteolysis ◽  
Histone H4 ◽  
In Vitro Activation

scholarly journals Protein kinase D1 phosphorylation of KAT7 enhances its protein stability and promotes replication licensing and cell proliferation

2020 ◽  
Vol 6 (1) ◽  
Author(s):  
Yao Liang ◽  
Yuanyuan Su ◽  
Chenzhong Xu ◽  
Na Zhang ◽  
Doudou Liu ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Dna Replication ◽  
Protein Kinase ◽  
Histone H4 ◽  
Replication Complex ◽  
Protein Kinase D1 ◽  
Defective Mutant ◽  
Histone H4 Acetylation

Abstract The histone acetyltransferase (HAT) KAT7/HBO1/MYST2 plays a crucial role in the pre-replication complex (pre-RC) formation, DNA replication and cell proliferation via acetylation of histone H4 and H3. In a search for protein kinase D1 (PKD1)-interacting proteins, we have identified KAT7 as a potential PKD1 substrate. We show that PKD1 directly interacts and phosphorylates KAT7 at Thr97 and Thr331 in vitro and in vivo. PKD1-mediated phosphorylation of KAT7 enhances its expression levels and stability by reducing its ubiquitination-mediated degradation. Significantly, the phospho-defective mutant KAT7-Thr97/331A attenuates histone H4 acetylation levels, MCM2/6 loading on the chromatin, DNA replication and cell proliferation. Similarly, PKD1 knockdown decreases, whereas the constitutive active mutant PKD1-CA increases histone H4 acetylation levels and MCM2/6 loading on the chromatin. Overall, these results suggest that PKD1-mediated phosphorylation of KAT7 may be required for pre-RC formation and DNA replication.

In vitro activation and substrates of recombinant, baculovirus expressed human protein kinase Cμ

FEBS Letters ◽  
1996 ◽  
Vol 381 (3) ◽  
pp. 183-187 ◽  
Author(s):  
Sabine Dieterich ◽  
Thomas Herget ◽  
Gisela Link ◽  
Heiner Böttinger ◽  
Klaus Pfizenmaier ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Recombinant Baculovirus ◽  
Human Protein ◽  
Human Protein Kinase ◽  
In Vitro Activation

In vitro activation of mouse skin protein kinase C by fatty acids and their hydroxylated metabolites

Lipids ◽  
1994 ◽  
Vol 29 (8) ◽  
pp. 547-553 ◽  
Author(s):  
Herng-Hsiang Lo ◽  
Gwendolyn A. Bartek ◽  
Susan M. Fischer
Keyword(s):  
Fatty Acids ◽  
Protein Kinase C ◽  
Protein Kinase ◽  
Mouse Skin ◽  
Hydroxylated Metabolites ◽  
Kinase C ◽  
In Vitro Activation ◽  
Skin Protein

scholarly journals The Archaeon Sulfolobus solfataricusContains a Membrane-Associated Protein Kinase Activity That Preferentially Phosphorylates Threonine Residues In Vitro

2000 ◽  
Vol 182 (12) ◽  
pp. 3452-3459 ◽  
Author(s):  
Brian H. Lower ◽  
Kenneth M. Bischoff ◽  
Peter J. Kennelly
Keyword(s):  
Protein Kinase ◽  
Molecular Mass ◽  
Kinase Activity ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Protein Kinase Activity ◽  
Histone H4 ◽  
Purine Nucleotides ◽  
Archaeal Protein

ABSTRACT The extreme acidothermophilic archaeon Sulfolobus solfataricus harbors a membrane-associated protein kinase activity. Its solubilization and stabilization required detergents, suggesting that this activity resides within an integral membrane protein. The archaeal protein kinase utilized purine nucleotides as phosphoryl donors in vitro. A noticeable preference for nucleotide triphosphates over nucleotide diphosphates and for adenyl nucleotides over the corresponding guanyl ones was observed. The molecular mass of the solubilized, partially purified enzyme was estimated to be ≈125 kDa by gel filtration chromatography. Catalytic activity resided in a polypeptide with an apparent molecular mass of ≈67 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Challenges with several exogenous substrates revealed the protein kinase to be relatively selective. Only casein, histone H4, reduced carboxyamidomethylated and maleylated lysozyme, and a peptide modeled after myosin light chains (KKRAARATSNVFA) were phosphorylated to appreciable levels in vitro. All of the aforementioned substrates were phosphorylated on threonine residues, while histone H4 was phosphorylated on serine as well. Substitution of serine for the phosphoacceptor threonine in the myosin light chain peptide produced a noticeably inferior substrate. The protein kinase underwent autophosphorylation on threonine and was relatively insensitive to a set of known inhibitors of “eukaryotic” protein kinases.

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