Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:1.13.12.7 (firefly luciferase)
 2,792 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

Ueda and Suzuki (1998. Biochim. Biophys. Acta. 1380:313-319; 1998. Biophys. J. 75:1052-1057) reported that myristic acid inhibited firefly luciferase in microM range in competition with luciferin, whereas anesthetics inhibited it in millimeter ranges noncompetitively with luciferin. Myristate increased, whereas anesthetics decreased, the thermal denaturation temperature. The present study showed that high pressure increased the steady-state light intensity of the halothane-doped firefly luciferase but decreased that of the myristate-doped firefly luciferase. The steady-state light intensity showed a maximum at 19.1 degrees C. At 19.1 degrees C, high pressure did not affect the light intensity in the absence of the inhibitors. In the presence of 0.5 mM halothane, however, 25 MPa pressure (maximum effect) increased the light intensity to 106.0% of the control without the inhibitor. In the presence of 2.5 microM myristate, 40 MPa pressure decreased the light intensity to 90.9% of the control. When the temperature was 25 degrees C in the absence of inhibitors, 40 MPa pressure increased the light intensity 119.2% of the ambient value. At 0.5 mM halothane, 40 MPa pressure further increased the light intensity to 106.1% above the control 40 MPa value. At 2.5 microM myristate, 40 MPa pressure decreased the light intensity to 90.1% of the control 40 MPa value. From the pressure dependence of the light intensity, the volume change DeltaV of the enzyme was estimated at 25 degrees C: 0.5 mM halothane increased DeltaV = +3.93 cm3 mol-1, whereas 2.5 microM myristate decreased DeltaV = -7.66 cm3 mol-1. Present results show that there are distinct differences between the specific and nonspecific ligands in their response to high pressure. Myristate, which competes with luciferin, decreased the protein volume and stabilized the conformation against thermal perturbation. Halothane, which does not compete with the substrate, increased the protein volume and destabilized the conformation.
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PMID:Does pressure antagonize anesthesia? Opposite effects on specific and nonspecific inhibitors of firefly luciferase. 987 61

The physiological effects of anesthetics have been ascribed to their interaction with hydrophobic sites within functionally relevant CNS proteins. Studies have shown that volatile anesthetics compete for luciferin binding to the hydrophobic substrate binding site within firefly luciferase and inhibit its activity (Franks, N. P., and Lieb, W. R. (1984) Nature 310, 599-601). To assess whether anesthetics also compete for ligand binding to a mammalian signal transduction protein, we investigated the interaction of the volatile anesthetic, halothane, with the Rho GDP dissociation inhibitor (RhoGDIalpha), which binds the geranylgeranyl moiety of GDP-bound Rho GTPases. Consistent with the existence of a discrete halothane binding site, the intrinsic tryptophan fluorescence of RhoGDIalpha was quenched by halothane (2-bromo-2-chloro-1,1,1-trifluoroethane) in a saturable, concentration-dependent manner. Bromine quenching of tryptophan fluorescence is short-range and W192 and W194 of the RhoGDIalpha are located within the geranylgeranyl binding pocket, suggesting that halothane binds within this region. Supporting this, N-acetyl-geranylgeranyl cysteine reversed tryptophan quenching by halothane. Short chain n-alcohols ( n < 6) also reversed tryptophan quenching, suggesting that RhoGDIalpha may also bind n-alkanols. Consistent with this, E193 was photolabeled by 3-azibutanol. This residue is located in the vicinity of, but outside, the geranylgeranyl chain binding pocket, suggesting that the alcohol binding site is distinct from that occupied by halothane. Supporting this, N-acetyl-geranylgeranyl cysteine enhanced E193 photolabeling by 3-azibutanol. Overall, the results suggest that halothane binds to a site within the geranylgeranyl chain binding pocket of RhoGDIalpha, whereas alcohols bind to a distal site that interacts allosterically with this pocket.
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PMID:Interaction of anesthetics with the Rho GTPase regulator Rho GDP dissociation inhibitor. 1870 20