Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.11.13 (protein kinase C)
 49,245 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Evidence is emerging that calcium-activated neutral proteinases (CANPs) not only participate in intracellular protein turnover but help to regulate the functional reorganization of cytoskeletal proteins in response to calcium and second-messenger stimulation. The high concentration of CANPs in certain neurons has suggested prominent roles for this proteolytic system in neuronal and synaptic function. In addition to acting directly on specific constituents of the cytoplasmic and membrane-associated cytoskeletal networks, CANP may amplify its effects by modulating the activities of protein kinase C and possibly other kinases and phosphatases by limited proteolysis. Given its suspected involvement at the cytoskeleton-membrane interface, calcium-mediated proteolysis is an example of a metabolic process which, if impaired, could provide a unifying basis for the slow progressive development of diverse structural and functional abnormalities within neurons. The multiplicity of mechanisms regulating its activity makes the CANP system a vulnerable target for disruption from various sources. A working hypothesis is advanced that down-regulation (inhibition) of neuronal calcium-mediated proteolysis in Alzheimer's disease is one critical and early step in the development of neurofibrillary degeneration and altered membrane cytoskeleton dynamics, which leads to membrane injury, accumulation of abnormal proteins, and synaptic dysfunction.
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PMID:Calcium-activated neutral proteinases as regulators of cellular function. Implications for Alzheimer's disease pathogenesis. 256 Sep

We show here a novel method for the in situ analysis of proteolyzed proteins in a cell. As a model, we focused on protein kinase C (PKC) beta, which is cleaved at a specific site between the catalytic and regulatory domains by calpain, the intracellular calcium-activated neutral proteinase. To detect proteolyzed PKC beta 'cleavage-site-directed antibodies', which specifically recognize the amino-terminal region of the catalytic fragment but do not cross-react with the unproteolyzed enzymes, were raised using synthetic peptide. The synthetic peptide used in this study was QGTKVPEEKTT, corresponding to the amino-terminal region of the catalytic fragment from human PKC beta generated by calpain. Rabbits were immunized with the synthetic peptide after conjugation with a carrier protein. Antibodies obtained reacted with the 46-kDa catalytic fragment of PKC beta, whereas they did not cross-react with unproteolyzed enzyme nor other fragments with different amino-termini. Thus, our antibody is specific to the amino-terminal sequence QGTKVPEEKTT, but does not recognize the same sequence located internally in native PKC beta. When human monoblast U937 cells were treated with calcium ionophore, the catalytic fragment of PKC beta was detected in the cytosol by immunoblotting with the antibody. However, this antibody did not bind unproteolyzed 80-kDa PKC beta, although this form was dominant in the cytosol of the calcium ionophore-treated cells. We could also detect comparable amounts of catalytic fragment in the calcium ionophore-treated cells by immunocytochemical staining with the same antibody. Our method was applied to examine the proteolysis of PKC beta in neutrophils stimulated with various reagents.
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PMID:Novel antibodies specific for proteolyzed forms of protein kinase C: production of anti-peptide antibodies available for in situ analysis of intracellular limited proteolysis. 844 81

The alpha isoform of protein kinase C (PKC alpha) is rapidly hydrolyzed by mM Ca(2+)-requiring calpain (calcium-activated neutral proteinase) under cell-free conditions (Shea et al, 1994, FEBS Lett. 350:223). In the present study, we demonstrate that this hydrolysis is inhibited by phosphatidyl serine, diacylglycerol, phosphatidyl choline, phosphatidyl inositol, and phosphatidic acid. With the exception of phosphatidic acid, these phospholipids did not directly inhibit calpain activity as evidenced by degradation of [14C]azocasein, suggesting that the nature of inhibition of calpain-mediated PKC alpha degradation is due to an effect of phospholipids on PKC alpha conformation. These findings suggest that m calpain-mediated PKC alpha hydrolysis may be specifically minimized at the plasma membrane, and leave open the possibility that such a mechanism exists in situ. In addition, the unique inhibition of calpain activity by phosphatidic acid suggests the existence of a specific mechanism by which this phospholipid regulates PKC alpha activity.
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PMID:Phospholipids inhibit proteolysis of protein kinase C alpha by mM calcium-requiring calpain. 878 23