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Query: UNIPROT:P06889 (Mol)
 630,302 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In low ionic strength buffer (5 mM Tris.HCl), the binding of [3H] nitrendipine to dihydropyridine calcium antagonist binding sites of mouse forebrain membranes is increased by both Na+ and Ca2+. Radiation inactivation was used to determine the target size of [3H]nitrendipine binding sites in 5 mM Tris.HCl buffer, in the presence and absence of these cations. After irradiation, [3H] nitrendipine binding in buffer with or without Na+ was diminished, due to a loss of binding sites and also to an increase in Kd. After accounting for radiation effects on the dissociation constant, the target size for the nitrendipine binding site in buffer was 160-170 kDa and was 170-180 kDa in the presence of sodium. In the presence of calcium ions, [3H]nitrendipine binding showed no radiation effects on Kd and yielded a target size of 150-170 kDa. These findings suggest, as in the case of opioid receptors, the presence of high molecular weight membrane components that modulate cation-induced alterations in radioligand binding to dihydropyridine binding sites.
Mol Pharmacol 1989 Aug
PMID:Radiation inactivation reveals discrete cation binding sites that modulate dihydropyridine binding sites. 254 88

The interaction of rat liver acetyl-CoA carboxylase with a 2',3'-dialdehyde derivative of ATP (oATP) has been studied. The degree of the enzyme inactivation has been found to depend on the oATP concentration and the incubation time. ATP was the only reaction substrate which provided protection from inactivation. Acetyl-CoA did not affect inactivation, while HCO3- accelerated the process. Ki values for oATP in the absence and the presence of HCO3- were 0.35 +/- 0.04 and 0.5 +/- 0.06 mM, and those of the modification constant (k) were 0.11 and 0.26 min-1, respectively. oATP completely inhibited the reaction of [14C]ADP in equilibrium ATP exchange, whereas produced actually no effect on [14C]acetyl-CoA equilibrium with malonyl-CoA exchange. Incorporation of about one equivalent of [3H]oATP per acetyl-CoA carboxylase subunit has been shown. No restoration of the modified enzyme activity has been observed in Tris or beta-mercaptoethanol containing buffers, and treatment with NaB[3H]4 has not led to 3H incorporation. The modification process involves elimination of the triphosphate chain of oATP. The results obtained indicate the affinity character of oATP-mediated modification of acetyl-CoA carboxylase. The reagent apparently interacts selectively with the epsilon-amino group of lysine in the ATP-binding site to form a morpholine-like structure.
Mol Biol (Mosk)
PMID:[Acetyl-CoA-carboxylase: modification of ATP-binding site of the active center by 2',3'-dialdehyde derivative of ATP]. 257 82

Rat liver lysosomes have been used to characterize further the effects of ATP on lysosomal stability during incubation at 37 degrees C at hypo-osmolarity. As previously reported, when the osmotically-supporting solute is the salt of a strong base (K+), ATP protects against lysis during incubation. However, if the osmotically-supporting solute is the salt of a weak base, e.g. Tris HCl or NH4Cl, ATP actually promotes lysis during incubation. Thus, ATP can exert destabilizing as well as protective effects on lysosomes. The destabilizing effect is eliminated by protonophores. The protective effect in the presence of potassium salts is not eliminated by protonophores. Moreover, when incubation is in the presence of a salt of a weak base, protonophores actually cause an ATP-dependent protective effect to be established. The destabilizing effect occurs at 37 degrees C, but not at 0 degrees C. The Mg++-dependence of the destabilizing effect was found to be similar to that found earlier for the ATP-dependent protective effect, insofar as only 1 mM MgCl2 in the presence of 1mM EDTA is sufficient for nearly maximal stimulation of both effects. The destabilizing effect may result from a H+ ion gradient across the lysosomal membrane which is maintained by the lysosomal ATP-dependent proton pump. The protective effect, on the other hand, does not depend on such a gradient being maintained; on the contrary, protonophores appear to act as enablers of the protective effect. The question that remains to be answered is: does the protective effect derive in some way from the same ATP-driven mechanism which constitutes the proton pump? Some possible answers to this question are considered.
Mol Cell Biochem 1989 Oct 31
PMID:Is the ATP-dependent protection of lysosomes against osmotic lysis a function of the lysosomal proton pump. 258 96

Casein kinase II from a virally-transformed macrophage cell line (RAW264) was purified by a sequential DEAE, Procion Red, phosvitin-Sepharose and heparin-Sepharose chromatography. With [tau-32P]GTP as a phosphate donor and casein as a substrate, the kinase was stimulated by polyamines and inhibited by heparin. The purified kinase had a specific activity of 1137 nmol/min/mg protein and exhibited three major protein bands of 40 K, 35 K, and 25 K. Under non-denaturing conditions in 50 mM Tris-50 mM NaCl the enzyme was eluted as a single peak with molecular weight of 110 K. Incubation of kinase in the presence of [tau-32P]GTP and Mg2+ resulted in phosphorylation of the 25 K protein band of the enzyme. In the presence of [tau-32P]GTP and Mg2+ the kinase was able to phosphorylate 55 K protein band in purified ornithine decarboxylase preparation from RAW264 cells and the rat-type II regulatory subunit of the cyclic AMP-dependent protein kinase.
Cell Mol Biol 1989
PMID:Characterization of casein kinase II from a virally transformed macrophage-like cell line, RAW264. 262 1

The specific DNA-binding protein FIS (factor for inversion stimulation), which stimulates site-specific DNA inversion by interaction with an enhancer sequence, was purified from an Escherichia coli strain overproducing the protein. FIS was crystallized at room temperature by microdialysis against 1.2 to 1.5 M-sodium/potassium phosphate containing 10 mM-Tris.HCl, 0.5 to 1 M-NaCl and 1 mM-NaN3 at pH 8.0 to 8.2. The crystals are stout prisms and suitable for X-ray diffraction study beyond 2.5 A resolution. They belong to the orthorhombic space group P2(1)2(1)2(1). The unit cell has dimensions a = 47.57(4) A, b = 51.13(4) A, c = 79.83(6) A and contains one FIS dimer in the asymmetric unit.
J Mol Biol 1989 Jul 05
PMID:Crystallization of the DNA-binding Escherichia coli protein FIS. 267 86

Hepatic estrogen receptors (ERs) of the female turtle, Chrysemys picta, when complexed with [3H]estradiol ([3H]E2), were shown to bind specifically to liver chromatin isolated from the same species. The binding of the [3H]E2 receptor complex to chromatin requires both the steroid ligand and the receptor protein. Maximal binding occurred within 60-70 min of incubation at 4 degrees C in a Tris buffer containing 0.1 M KCl. The binding of the [3H]E2 receptor complex to intact chromatin was saturable, whereas the binding to turtle or calf thymus DNA remained linear. Scatchard analyses revealed more estrogen receptor binding sites on hepatic chromatin isolated from female turtles than that prepared from the males (binding capacities: female chromatin = 67.9 +/- 6.8 fmol/mg DNA equivalent; male chromatin = 28.5 +/- 2.5 fmol/mg DNA equivalent). Furthermore, the [3H]E2 receptor complex was bound with a higher affinity to female chromatin than to male chromatin (association constants: female chromatin = 11.7 +/- 2.7 X 10(10) M-1; male chromatin = 2.5 +/- 0.7 X 10(10) M-1). In contrast to turtle hepatic [3H]E2 receptors, ERs in rat liver or mouse uterine cytosol exhibited little binding affinity for hepatic chromatin isolated from the turtle. Tissue specificity was demonstrated in the interaction of the [3H]E2 receptor complex and chromatin; high affinity, saturable binding of the [3H]E2 receptor complex was only observed on chromatin isolated from the liver but not on those prepared from the heart, kidney and muscle. A 3- to 4-fold increase in the number of hepatic chromatin [3H]E2 receptor binding sites was observed in 21-day ovariectomized or hypophysectomized female (capacities = 209.3 +/- 6.1 and 270 +/- 10.1 fmol/mg DNA equivalent, respectively). It is postulated that [3H]E2 receptor binding sites on the chromatin of intact females are partially 'masked', and removal of a gonadal and/or pituitary factor(s) unveils additional binding sites on the female chromatin. This paper is first to report the presence of high affinity, species- and tissue-specific acceptor sites on the liver chromatin of a reptilian species. The fact that the levels and properties of these acceptor sites are dependent on the sex and hormonal state of the animal suggests that they may play a role in the regulation of hepatic estrogen responsiveness and vitellogenesis in this species.
Mol Cell Endocrinol 1989 Jan
PMID:Nuclear acceptor sites for estrogen-receptor complexes in the liver of the turtle, Chrysemys picta. I. Sexual differences, species specificity and hormonal dependency. 274 16

We have measured the contribution of the alkaline Bohr effect of the C-terminal histidine residues of the beta-chains of haemoglobin A by comparing haemoglobin A with haemoglobin Cowtown in which those histidine residues are replaced by leucine. Oxygenation of a stripped 2.5 mM (haem) solution of haemoglobin A yielded 0.19 H+/haem, while oxygenation of a similar solution of haemoglobin Cowtown produced no change of pH. Oxygen equilibria measured at 60 microM-haem in 0.1 M-Hepes buffer gave an alkaline Bohr effect of -0.21 H+/haem for haemoglobin A and only -0.01 H+/haem for haemoglobin Cowtown, even though its Hill's coefficient was greater than 2 throughout the pH range studied. These results prove that the chloride-independent part of the alkaline Bohr effect is due to the C-terminal histidine residues of the beta-chains. Oxygen equilibria measured in 0.095 M-bis-Tris buffers with minimal chloride or with 0.1 M-chloride showed the contribution of those histidine residues to the alkaline Bohr effect to be about 0.2 H+/haem, independent of chloride concentration. Determination of the individual Adair coefficients in the three different buffers indicated that pH and chloride tend to have their greatest effects at the second or third steps of oxygenation when the change of quaternary structure is most likely to occur; between pH 7 and 9, the fourth Adair coefficient is only very slightly affected by pH and not significantly by chloride.
J Mol Biol 1987 May 20
PMID:Influence of anions and protons on the Adair coefficients of haemoglobins A and Cowtown (His HC3(146) beta----Leu). 282 Dec 77

Electropherograms of Neurospora crassa homogenates showed a polypeptide with a mobility slightly lower than that of a standard sample of clathrin (from bovine brain). Subcellular fractionation of the homogenate resulted in a 20-fold enrichment of the putative N. crassa clathrin in the microsomal fraction. Further fractionation of the microsomal fraction by glass bead permeation chromatography yielded a fraction enriched about 150-fold relative to the homogenate. Coated vesicles (42.5 +/- 2.5 nm diameter) were found in this preparation by electron microscopy of negatively stained specimens. Ribosomes were virtually absent from this sample. N. crassa clathrin remained associated with the coated vesicles after repeated centrifugation and homogenization steps, even in the presence of 0.4 M-NaCl, but was released by treatment with Tris buffer pH 8.5. However the polypeptide was again sedimentable after dialysis against Mes buffer pH 6.5. Under the electron microscope this sediment resembled the empty coats of higher eukaryotes. The results taken together indicate that a clathrin-like protein occurs in wild type cells of N. crassa.
Mol Cell Biochem 1987 Sep
PMID:Clathrin coated vesicles in Neurospora crassa. 289 27

The MgATPase activity of the rabbit skeletal myosin subfragment 1 (S1), in the steady state, was measured by means of the intrinsic fluorescence of tryptophan. This technique gave results similar to those obtained by other methods (linked or radioactive assays). The activity was measured under conditions that effect the monomer/dimer ratio. It is shown that there is a close correlation between MgATPase activity and the proportion of dimer. At 20 degrees C, for pH 6.9 to 8.1 and for [KCl] less than or equal to 1 M, the observed activity (kobs) can be linearly related to the proportion of dimer (Ed/Eo) by: kobs(s-1) = 0.016-7 X 10(-3)[KCl] + 0.031(Ed/Eo), where [KCl] is expressed in M. We deduce that, at 20 degrees C and for [KCl] = 0 M, the activity of the monomer is kmobs = 0.016 s-1 (Ed/Eo = 0) and that of the dimer kdobs = 0.047 s-1 (Ed/Eo = 1), i.e. a ratio kdobs/kmobs approximately equal to 3. Beyond pH approximately equal to 8.3, the activities of both the monomer and the dimer increased steeply with increasing pH value. In the standard conditions (pH 8.0, [KCl] = 0 to 100 mM), S1 is mainly in the form of a dimer, and such conditions are not appropriate for study of the S1 monomer. For studying the pure monomer, the conditions required at 20 degrees C and in bis-Tris-propane are: S1 concentration approximately equal to 0.2 mg/ml, pH 6.9 to 7.8, [KCl] approximately equal to 300 mM. For studying the pure dimer, the conditions required are: S1 concentration greater than or equal to 0.2 mg/ml, pH 7.8 to 8.1 and [KCl] approximately equal to 0. In both cases the MgATP concentration is about 50 microM. Finally, if great care is taken concerning the age of the S1 solutions and the evaluation of the proportion of dimer, the values of kobs are extremely precise: the uncertainty regarding the values of kobs, as determined by means of intrinsic fluorescence, does not exceed +/- 0.001 s-1. Beyond this error bar conditions are uncontrolled.
J Mol Biol 1986 Sep 20
PMID:MgATPase activity of myosin subfragment 1. The dimer is more active than the monomer. 294 83

Oligomycin-sensitive particulate ATPase (MB ATPase) from L. donovani promastigotes was solubilized by chloroform treatment. Polyacrylamide gel electrophoresis revealed several protein bands, with the major one possessing ATPase activity. The solubilized enzyme had Mg2+-ATPase and Ca2+-ATPase but no K+-dependent alkaline phosphatase activity. The Mg2+-ATPase activity was stimulated by monovalent cations and was not sensitive to oligomycin. Hence it is referred to as F1 ATPase. It had optimum activity at pH 7.6 and 30 degrees C. The Arrhenius plot for MB ATPase was biphasic with activation energies (Ea) of 16.2 and 3.4 kcal mol-1, while F1 ATPase exhibited a linear plot with Ea = 10.1 kcal mol-1. Lineweaver-Burk plots were biphasic with Km values of 0.17 and 1.25 mM for MB ATPase and 0.18 and 1.33 mM for F1 ATPase. The enzyme could be preserved at -15 degrees C in Tris-SO2-(4)-EDTA-ATP-glycerol (t1/2 = 20 days).
Mol Biochem Parasitol 1988 Jun
PMID:Solubilization and kinetic characterization of mitochondrial adenosine triphosphatase from Leishmania donovani promastigotes. 297 May 89


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