Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: EC:3.1.4.3 (phospholipase C)
 18,461 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Transepithelial phosphate (Pi) fluxes were determined in primary monolayer cultures of the winter flounder proximal tubule in Ussing chambers. Net Pi secretion, peritubular (P)-to-lumen (L) net flux (Jnet = 17.0 +/- 3.77 nmol.cm-2.h-1), was strongly stimulated by lowering peritubular pH to 6.5 (pHP 6.5 vs. pHL 7.5) compared with control tissues at pH 7.5 (pHP 7.5 vs. pHL 7.5) where net reabsorption (L-to-P Jnet = 1.10 +/- 0.36 nmol.cm-2.h-1) occurred. The stimulation of net secretion by pHP 6.5 was inhibited to 27% of control by 200 microM amiloride. The imposition of a reversed pH gradient (pHL 6.5 vs. pHP 7.5) did not stimulate Pi secretion. Preincubation with 0.5 U/ml phospholipase C caused a nearly fivefold stimulation of Pi secretion compared with the untreated controls. H-7 (100 microM), a protein kinase inhibitor, caused a 2.5-fold reduction in phorbol ester (PE)-induced stimulation of Pi secretion. H-7 also significantly inhibited Pi secretion (50% reduction) when peritubular pH was lowered to 6.5. PE-induced net Pi secretion was significantly lower when luminal pH was lowered to 5.5 compared with controls where luminal pH was kept at 7.5. Amiloride (200 microM) significantly inhibited the PE-induced net Pi secretion in the presence of luminal pH 7.5 but had no significant effect at luminal pH 5.5. Replacement of luminal NaCl with LiCl or isosmolar mannitol significantly reduced net phosphate secretion under the conditions of lowered peritubular pH. Net Pi secretion was also dependent on the maintenance of a cellular Na+ gradient, since 100 microM ouabain inhibited PE-induced net Pi secretion.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:Effects of pH on phosphate transport by flounder renal tubule primary cultures. 184 76

Activation of the Na+/H+ antiport mechanism was studied in human neutrophils by monitoring intracellular pH with a carboxyfluorescein derivative. N-formyl-methionyl-leucyl-phenylalanine (FMLP) and phospholipase C (PLC) induced biphasic pH changes. Amiloride, which inhibits the antiport, completely blocked alkalinization but enhanced acidification. Polymyxin B, which inhibits protein kinase C, only blocked alkalinization. Activation with phorbol 12-myristate 13-acetate (PMA) led to alkalinization only; this was inhibited by amiloride or polymyxin B. Thus, during polymorphonuclear leukocyte (PMN) activation, intracellular alkalinization appears to be mediated by an amiloride-sensitive Na+/H+ antiport. Antiport activity can also be blocked indirectly by inhibition of protein kinase C activity. Early intracellular acidification does not appear to require kinase activity but is observed when phospholipids are remodeled with PLC. The antiport was also activatable by hypertonic buffered media. This response did not appear to be mediated by protein kinase C because it was unaffected by polymyxin B. Finally, superoxide generation was investigated. It is affected by, but not soley controlled by, either antiport or protein kinase C activity.
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PMID:Intracellular pH changes during neutrophil activation: Na+/H+ antiport. 302 17

1. The aim of the present study was to validate the Cytosensor microphysiometer, a novel system that measures the extracellular acidification rate as a reliable index of the integrated functional response to receptor activation, as a method for studying NK3 receptor pharmacology, and then to use this system to assess the functional activity of novel compounds at this receptor. 2. The selective NK3 agonist senktide caused reproducible, concentration-related increases in acidification ratein CHO-NK3 cells, with a pEC50 value of 8.72+/-0.11 (n=15). [Beta-Ala8]NKA(4-10), the selective NK2 agonist, elicited a much weaker response (pEC50=6.68+/-0.08, n=4), while the NK1-selective agonist substance P methylester only caused a very weak response at concentrations > or =3 microM (n=2). The rank order of potency for the endogenous tachykinins NKB>NKA>substance P (n=3) confirmed the response was mediated by the NK3 receptor. Moreover, the actual potencies obtained were consistent with affinities measured in radioligand binding studies. 3. The novel compounds PD156319-121 (0.3-1 microM), PD161182 (10-300 nM), PD168001 (10-100 nM) and PD168073 (10-100 nM) all acted as surmountable antagonists of the senktide-induced acidification response, with pA2 values of 7.49, 8.67, 9.17 and 9.25 respectively (n=3-5). In comparison the known NK3 antagonist SR142801 (10-100 nM) had a pA2 value of 8.83 (n=8) for the interaction with senktide. Again, these values are consistent with the radioligand binding data. 4. Amiloride (1 mM) inhibited the senktide-induced acidification response by 68.3+/-3.3 (n=4), indicating that the Na+/H+ antiporter plays an important role in this response, and this is consistent with the importance of this antiporter in other acidification responses. 5. Inhibition of protein kinase C with staurosporine (0.1 microM), or depletion of the intracellular Ca2+ stores with thapsigargin (1 microM), both resulted in a reduction in the maximum response to senktide (63.3+/-1.7 and 68.9+/-3.2% respectively, n=3-5), and co-application of these inhibitors abolished the response (n=3). This strongly suggested that the NK3 receptor was coupling via phospholipase C (PLC), as would be expected, although this could not be confirmed by the use of the putative PLC/PLA2 inhibitor U73122. 6. In conclusion, we have demonstrated the utility of the Cytosensor in the characterization of functional responses to agonists, and assessment of the affinities of antagonists in CHO cells expressing the human NK3, and have shown that our series of novel compounds are non-peptide NK3 antagonists of high affinity, as exemplified by PD168073.
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PMID:Activation of the cloned human NK3 receptor in Chinese Hamster Ovary cells characterized by the cellular acidification response using the Cytosensor microphysiometer. 983 12