Redox regulation of Ca2+/calmodulin-dependent protein kinase IV via oxidation of its active-site cysteine residue

2019 ◽  
Vol 130 ◽  
pp. 99-106 ◽  
Author(s):  
Tsuyoshi Takata ◽  
Jun Kimura ◽  
Hideshi Ihara ◽  
Naoya Hatano ◽  
Yukihiro Tsuchiya ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Active Site ◽  
Redox Regulation ◽  
Cysteine Residue ◽  
Dependent Protein Kinase ◽  
Active Site Cysteine ◽  
Dependent Protein ◽  
Active Site Cysteine Residue

scholarly journals Inactivation of Ca2+/calmodulin-dependent protein kinase I byS-glutathionylation of the active-site cysteine residue

FEBS Letters ◽  
2010 ◽  
Vol 584 (11) ◽  
pp. 2478-2484 ◽  
Author(s):  
Toshie Kambe ◽  
Tao Song ◽  
Tsuyoshi Takata ◽  
Naoya Hatano ◽  
Yoshiaki Miyamoto ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Active Site ◽  
Cysteine Residue ◽  
Dependent Protein Kinase ◽  
Active Site Cysteine ◽  
Dependent Protein ◽  
Active Site Cysteine Residue

Reactive sulfur species inactivate Ca2+/calmodulin-dependent protein kinase IV via S-polysulfidation of its active-site cysteine residue

2017 ◽  
Vol 474 (15) ◽  
pp. 2547-2562 ◽  
Author(s):  
Tsuyoshi Takata ◽  
Hideshi Ihara ◽  
Naoya Hatano ◽  
Yukihiro Tsuchiya ◽  
Takaaki Akaike ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Er Stress ◽  
Active Site ◽  
Cysteine Residue ◽  
Dependent Protein Kinase ◽  
Sulfur Species ◽  
Active Site Cysteine ◽  
Dependent Protein ◽  
Reactive Sulfur Species

Reactive sulfur species (RSS) modulate protein functions via S-polysulfidation of reactive Cys residues. Here, we report that Ca2+/calmodulin (CaM)-dependent protein kinase IV (CaMKIV) was reversibly inactivated by RSS via polysulfidation of the active-site Cys residue. CaMKIV is phosphorylated at Thr196 by its upstream CaMK kinase (CaMKK), resulting in the induction of its full activity. In vitro incubation of CaMKIV with the exogenous RSS donors Na2Sn (n = 2–4) resulted in dose-dependent inhibition of the CaMKK-induced phospho-Thr196 and consequent inactivation of the enzyme activity. Conversely, mutated CaMKIV (C198V) was refractory to the Na2Sn-induced enzyme inhibition. A biotin-polyethylene glycol-conjugated maleimide capture assay revealed that Cys198 in CaMKIV represents a target for S-polysulfidation. Furthermore, phosho-Thr196 and CaMKIV activity were inhibited by incubation with cysteine hydropersulfide, a newly identified RSS that is generated from cystine by cystathionine-γ-lyase. In transfected cells expressing CaMKIV, ionomycin-induced CaMKIV phosphorylation at Thr196 was decreased upon treatment with either Na2S4 or the endoplasmic reticulum (ER) stress inducer thapsigargin, whereas cells expressing mutant CaMKIV (C198V) were resistant to this treatment. In addition, the ionomycin-induced phospho-Thr196 of endogenous CaMKIV was also inhibited by treatment either with Na2S4 or thapsigargin in Jurkat T lymphocytes. Taken together, these data define a novel signaling function for intracellular RSS in inhibiting CaMKIV activity via S-polysulfidation of its Cys198 during the response to ER stress.

Substrate-mediated channeling of a chemical reagent to the active site of cAMP-dependent protein kinase

FEBS Letters ◽  
1981 ◽  
Vol 130 (1) ◽  
pp. 127-132 ◽  
Author(s):  
Juan S. Jiménez ◽  
Abraham Kupfer ◽  
Philip Gottlieb ◽  
Shmuel Shaltiel
Keyword(s):  
Protein Kinase ◽  
Active Site ◽  
Chemical Reagent ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

scholarly journals Insights into the phosphoryl transfer catalyzed by cAMP-dependent protein kinase

2014 ◽  
Vol 70 (a1) ◽  
pp. C449-C449
Author(s):  
Oksana Gerlits ◽  
Amit Das ◽  
Jianhui Tian ◽  
Malik Keshwani ◽  
Susan Taylor ◽  
...  
Keyword(s):  
Protein Kinase ◽  
Protein Kinases ◽  
Active Site ◽  
Phosphate Group ◽  
Nucleoside Triphosphate ◽  
Phosphoryl Transfer ◽  
Hydrogen Atoms ◽  
Dependent Protein Kinase ◽  
Camp Dependent Protein Kinase ◽  
Dependent Protein

Protein kinases are involved in a number of cell signaling pathways. They catalyze phosphorylation of proteins and regulate the majority of cellular processes (such as growth, differentiation, lipid metabolism, regulation of sugar, nucleic acid synthesis, etc.). Chemically, protein kinases covalently transfer the gamma-phosphate group of a nucleoside triphosphate (e.g. ATP) to a hydroxyl group of a Ser, Thr or Tyr residue of substrate protein or peptide. The reaction involves moving hydrogen atoms between the enzyme, substrate and nucleoside. The unanswered question is whether the proton transfer from the Ser residue happens before the phosphoryl transfer using the general acid-base catalyst, Asp166, or after the reaction went through the transition state by directly protonating the phosphate group. To address this key question about the phosphoryl transfer, we determined a number of X-ray structures of ternary complexes of catalytic subunit of cAMP-dependent protein kinase (PKAc) with various substrates, nucleotides and cofactors. Importantly, we were able to trap and mimic the initial (Michaelis complex) and final (product complex) stages of the reaction. The results demonstrate that Mg2+, Ca2+, Sr2+, and Ba2+ metal ions bind to the active site and facilitate the reaction to produce ADP and a phosphorylated peptide. The study also revealed that metal-free PKAc can facilitate the phosphoryl transfer reaction; a result that was confirmed with single turnover enzyme kinetics measurements. Comparison of the product and the pseudo-Michaelis complex structures, in conjunction with molecular dynamics simulations, reveals conformational, coordination, and hydrogen bonding changes that help further our understanding of the mechanism, roles of metals, and active site residues involved in PKAc activity.

scholarly journals The location of the active-site histidine residue in the primary sequence of papain

1968 ◽  
Vol 108 (5) ◽  
pp. 861-866 ◽  
Author(s):  
S. S. Husain ◽  
G. Lowe
Keyword(s):  
Amino Acid ◽  
Sodium Borohydride ◽  
Active Site ◽  
Iodoacetic Acid ◽  
Cysteine Residue ◽  
Histidine Residue ◽  
Primary Sequence ◽  
Active Site Cysteine ◽  
Active Site Histidine ◽  
Active Site Cysteine Residue

Papain that had been irreversibly inhibited with 1,3-dibromo[2−14C]acetone was reduced with sodium borohydride and carboxymethylated with iodoacetic acid. After digestion with trypsin and α-chymotrypsin the radioactive peptides were purified chromatographically. Their amino acid composition indicated that cysteine-25 and histidine-106 were cross-linked. Since cysteine-25 is known to be the active-site cysteine residue, histidine-106 must be the active-site histidine residue.

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