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D M Quinn

Publications and source records attributed to D M Quinn.

At least 55 records · Page 3Linked to original sources
PubMed1986

Phenyl-n-butylborinic acid is a potent transition state analog inhibitor of lipolytic enzymes.

The cholesterol esterase and lipoprotein lipase catalyzed hydrolyses of the water-soluble substrate p-nitrophenyl butyrate are competitively inhibited by butaneboronic acid and phenylboronic acid. Phenyl-n-butylborinic acid has been synthesized and characterized as an ultrapotent transition state analog inhibitor: Ki = 2.9 +/- 0.6 nM and 1.7 +/- 0.3 microM for the cholesterol esterase and lipoprotein lipase reactions, respectively. These results are interpreted in terms of transition state structure and stabilization.

Animals↗
PubMed1986

Prevalence of psychoactive medication in children and adolescents.

Prevalence studies in Psychopharmacology are few and have many methodological flaws. This study examines the prevalence of psychoactive drugs in children and adolescents. Contrary to other studies, central nervous system stimulants are not the most widely prescribed psychoactive drugs in childhood and adolescence, but rather, minor tranquilizers, sedatives and hypnotics are the most widely prescribed psychoactive drugs. While this study overcomes some of the flaws of previous studies further studies regarding the prevalence of use of psychoactive drugs in childhood and adolescence are recommended.

Adolescent↗
PubMed1985

Acylenzyme mechanism and solvent isotope effects for cholesterol esterase-catalyzed hydrolysis of p-nitrophenyl butyrate.

The mechanism of cholesterol esterase- (carboxylic ester hydrolase, EC 8.1.1.1) catalyzed hydrolysis of the water-soluble ester p-nitrophenyl butyrate has been characterized for commercially available preparations from bovine and porcine pancreas and for a purified preparation from porcine pancreas. Kinetic evidence for an acylenzyme mechanism is provided by experiments wherein the butyryl enzyme is trapped by MeOH, EtOH or n-BuOH. For the last alcohol the transacylation product n-butyl n-butyrate was characterized by GC-mass spectrometry. Solvent isotope effects have been measured for Vmax/Km, which is the rate constant for acylation, and for Vmax, which monitors rate-determining deacylation. Isotope effects of 1.5-3 on these rate constants indicate that both steps of the acylenzyme mechanism for cholesterol esterase catalysis involve transition states that are stabilized by general acid-base proton bridges.

Alcohols↗
PubMed1985

Solvent isotope effects for lipoprotein lipase catalyzed hydrolysis of water-soluble p-nitrophenyl esters.

Solvent deuterium isotope effects on the rates of lipoprotein lipase (LpL) catalyzed hydrolysis of the water-soluble esters p-nitrophenyl acetate (PNPA) and p-nitrophenyl butyrate (PNPB) have been measured and fall in the range 1.5-2.2. The isotope effects are independent of substrate concentration, LpL stability, and reaction temperature and hence are effects on chemical catalysis and not due to a medium effect of D2O on LpL stability and/or conformation. pL (L = H or D) vs. rate profiles for the Vmax/Km of LpL-catalyzed hydrolysis of PNPB increase sigmoidally with increasing pL. Least-squares analysis of the profiles gives pKaH2O = 7.10 +/- 0.01, pKaD2O = 7.795 +/- 0.007, and a solvent isotope effect on limiting velocity at high pL of 1.97 +/- 0.03. Because the pL-rate profiles are for the Vmax/Km of hydrolysis of a water-soluble substrate, the measured pKa's are intrinsic acid-base ionization constants for a catalytically involved LpL active-site amino acid side chain. Benzeneboronic acid, a potent inhibitor of LpL-catalyzed hydrolysis of triacylglycerols [Vainio, P., Virtanen, J. A., & Kinnunen, P. K. J. (1982) Biochim. Biophys. Acta 711, 386-390], inhibits LpL-catalyzed hydrolysis of PNPB, with Ki = 6.9 microM at pH 7.36, 25 degrees C. This result and the solvent isotope effects for LpL-catalyzed hydrolysis of water-soluble esters are interpreted in terms of a proton transfer mechanism that is similar in many respects to that of the serine proteases.

Animals↗
PubMed1985

Diethyl-p-nitrophenyl phosphate: an active site titrant for lipoprotein lipase.

Diethyl-p-nitrophenyl phosphate is an active site-directed irreversible inhibitor of bovine milk lipoprotein lipase catalyzed hydrolysis of the water-soluble substrate, p-nitrophenyl butyrate. Interaction of lipoprotein lipase and the inhibitor in the absence of substrate gives a biphasic kinetics profile, which is consistent with rapid formation of a phosphoryl-lipoprotein lipase intermediate which hydrolyzes slowly. The magnitude of the absorbance increase accompanying formation of the intermediate provides an analytical method for determining lipoprotein lipase active site concentration.

Animals↗
PubMed1985

The effectiveness, in vitro, of miconazole and ketoconazole combined with tissue conditioners in inhibiting the growth of Candida albicans.

Two recent antifungal agents, miconazole and ketoconazole, were combined with three tissue conditioners and tested in vitro for their effects on the growth of Candida albicans. Studies for comparison were carried out using the earlier antifungal agents, amphotericin B and nystatin. Miconazole and ketoconazole were as effective as nystatin in completely inhibiting the growth of Candida albicans. The ineffectiveness of amphotericin B when combined with tissue conditioners as an antifungal agent was confirmed. (See Addendum p.181.).

Acrylic Resins↗
PubMed1983

Lipoprotein lipase-catalyzed hydrolysis of phosphatidylcholine of guinea pig very low density lipoproteins and discoidal complexes of phospholipid and apolipoprotein: effect of apolipoprotein C-II on the catalytic mechanism.

To elucidate the mechanism by which apolipoprotein C-II (apoC-II) enhances the activity of lipoprotein lipase (LpL), discoidal phospholipid complexes were prepared with apoC-III and di[(14)C]palmitoyl phosphatidylcholine (DPPC) and containing various amounts of apoC-II. The rate of DPPC hydrolysis catalyzed by purified bovine milk LpL was determined on the isolated complexes. The rate of hydrolysis was optimal at pH 8.0. Analysis of enzyme kinetic data over a range of phospholipid concentrations revealed that the major effect of apoC-II was to increase the maximal velocity (V(max)) some 50-fold with a limited effect on the Michaelis constant (K(m)). V(max) of the apoC-III complex containing no apoC-II was 9.2 nmol/min per mg LpL vs. 482 nmol/min per mg LpL for the complex containing only apoC-II. The effect of apoC-II on enzyme kinetic parameters for LpL-catalyzed hydrolysis of DPPC complexes was compared to that on the parameters for hydrolysis of DPPC and trioleoylglycerol incorporated into guinea pig very low density lipoproteins (VLDL(p)) which lack the equivalent of human apoC-II. Tri[(3)H]oleoylglycerol-labeled VLDL(p) were obtained by perfusion of guinea pig liver with [(3)H]oleic acid. Di[(14)C]palmitoyl phosphatidylcholine was incorporated into the VLDL(p) by incubation of VLDL(p) with sonicated vesicles of di[(14)C]palmitoyl phosphatidylcholine and purified bovine liver phosphatidylcholine exchange protein. The rates of LpL-catalyzed hydrolysis of trioleoylglycerol and DPPC were determined at pH 7.4 and 8.5 in the presence and absence of apoC-II. In the presence of apoC-II, the V(max) for DPPC hydrolysis in guinea pig VLDL(p) increased at both pH 7.4 and pH 8.5 (2.4- and 3.2-fold, respectively); the value of K(m) did not change at either pH (0.23 mm). On the other hand, the kinetic value of K(m) for triacylglycerol hydrolysis in the presence of apoC-II decreased at both pH 7.4 (3.05 vs. 0.54 mm) and pH 8.5 (2.73 vs. 0.62 mm). These kinetic studies suggest that apoC-II enhances phospholipid hydrolysis by LpL in apoC-III-DPPC discoidal complexes and VLDL(p) mainly by increasing the V(max) of the enzyme for the substrates, whereas the activator protein primarily causes a decrease in the apparent K(m) for triacylglycerol hydrolysis.-Shirai, K., T. J. Fitzharris, M. Shinomiya, H. G. Muntz, J. A. K. Harmony, R. L. Jackson and D. M. Quinn. Lipoprotein lipase-catalyzed hydrolysis of phosphatidylcholine of guinea pig very low density lipoproteins and discoidal complexes of phospholipid and apolipoprotein: effect of apolipoprotein C-II on the catalytic mechanism.

Animals↗
PubMed1982

Lipoprotein lipase catalyzed hydrolysis of water-soluble p-nitrophenyl esters. Inhibition by apolipoprotein C-II.

Bovine milk lipoprotein lipase (LpL) catalyzes the hydrolysis of the water-soluble esters p-nitrophenyl acetate (PNPA) and p-nitrophenyl butyrate (PNPB). The same protein and same active site are involved in hydrolysis of water-soluble p-nitrophenyl esters and emulsified trioleoylglycerol since (a) trioleoylglycerol hydrolysis and PNPB hydrolysis activities coelute from the heparin-Sepharose affinity column used to purify LpL and (b) LpL-catalyzed hydrolyses of trioleoylglycerol and PNPB are inhibited to equal extents by phenylmethanesulfonyl fluoride. The effect of apolipoprotein C-II (apoC-II) on the LpL-catalyzed hydrolysis of PNPA and PNPB has been determined. ApoC-II inhibits hydrolysis of both esters, with a maximum extent of inhibition of 70-90%. Inhibition of the LpL-catalyzed hydrolysis of PNPB is specific for apoC-II, since apolipoproteins A-I, C-I, and C-III-2 have little effect on this reaction, and is partial noncompetitive in form. KI values for apoC-II inhibition of the LpL-catalyzed hydrolysis of PNPA and PNPB are in the range 0.26-0.83 microM. The effect of apoC-II on the temperature dependences of LpL-catalyzed hydrolysis of both esters and on NaCl inhibition of LpL-catalyzed PNPB hydrolysis is consistent with a change in rate-determining step with LpL and apoC-II interact. These results indicate not only that there is an interaction between apoC-II and LpL in aqueous solution in the absence of a lipid interface but also that this interaction conformationally modulates the active site of the enzyme.

Animals↗
PubMed1982

Reciprocal effect of apolipoprotein C-II on the lipoprotein lipase-catalyzed hydrolysis of p-nitrophenyl butyrate and trioleoylglycerol.

Interaction of purified bovine milk lipoprotein lipase (LpL) with sonicated vesicles of dipalmitoyl phosphatidylcholine in the gel phase is associated with an increase in the rate of the LpL-catalyzed hydrolysis of p-nitrophenyl butyrate. There is a 6-fold increase in Vmax. Apolipoprotein C-II, the activator protein for LpL, inhibits the LpL-catalyzed hydrolysis of p-nitrophenyl butyrate. With 0.5 mol % tri[14C]oleoylglycerol present in the dipalmitoyl phosphatidylcholine vesicles and in the presence of 20 mM Ca2+, the rate of p-nitrophenyl butyrate hydrolysis is decreased reciprocally compared to trioleoylglycerol hydrolysis and is dependent on apolipoprotein C-II. These results suggest that apolipoprotein C-II enhances the activity of LpL by increasing the affinity of the active site of LpL for triacylglycerol.

Animals↗
PubMed1982

Anisotropic motion of the steroid ring system of cholesteryl esters. Calculation of carbon-13 nuclear magnetic resonance relaxation times and nuclear Overhauser enhancements and comparison with experiment.

A quantitative model for the molecular dynamics of the steroid fused ring system of cholesteryl esters is discussed. The model describes rotational diffusive motion of the steroid rings as that of axially symmetric prolate ellipsoids and is used to generate predictions for the dependence of carbon-13 nuclear magnetic resonance (13C NMR) line widths, spin-lattice relaxation times, and nuclear Overhauser enhancements of C3 and C6 of cholesteryl esters on the correlation times for rotation about the symmetry axis and about the nonunique axes of the ellipse by using the spectral density functions developed by Woessner [Woessner, D.E. (1962) J. Chem. Phys. 37, 647-654]. The predictions are used to calculate correlation times for motion of the steroid rings of isotropic liquid-phase cholesteryl linoleate and cholesteryl oleate from NMR spectra at magnetic field strengths of 2.35 and 6.34 T and at various temperatures. Such calculations characterize steroid ring motion of cholesteryl esters as highly anisotropic, with motion about the symmetry axis 40-130 times faster than about the nonunique axes. The fact that the line width of the C6 resonance is consistently narrower than that of C3 is attributed to the high rotational anisotropy of cholesteryl esters and to the inclination of the C6-H internuclear vector at an angle with respect to the molecular symmetry axis that is very close to the "magic angle".

Cholesterol Esters↗
PubMed1982

Kinetics of plasma protein-catalyzed exchange of phosphatidylcholine and cholesteryl ester between plasma lipoproteins.

A lipid transfer complex (LTC) isolated from human plasma catalyzes equimolar exchange of cholesteryl ester and phosphatidylcholine between low density (LDL) and high density (HDL) plasma lipoproteins. Activation parameters for LTC-catalyzed exchange of neutral and polar lipid are equal and are not influenced by the degree of purity of the catalyst. Activation parameters for exchange of both cholesteryl ester and phosphatidylcholine are influenced by the extent of saturation of phosphatidylcholine fatty acyl groups. The activation parameters also depend on the amount of HDL present in the assay. The flux rates of lipid exchange depend on the concentration of both LDL and HDL. At constant HDL concentration, flux rates become independent of LDL concentration when the ratio of [LDL]:[HDL] exceeds 9:1 (based on cholesteryl ester); at constant LDL concentration, facilitated LDL,HDL lipid exchange is inhibited at high HDL concentration, suggesting preferential HDL,HDL exchange. Analysis of the dependence of initial lipid exchange rate on LDL concentration at two constant HDL concentrations suggests that, in the reaction pathway, LTC mediates a productive collision (ternary complex) between LDL and HDL. A kinetic mechanism consistent with the data is one in which lipid exchange occurs in a ternary complex consisting of LTC, HDL and LDL. At low HDL concentration, this complex is formed by a random sequential route; at high HDL concentration, the mechanism is ordered sequential since the reactants are an LTC-HDL complex and LDL.

Carrier Proteins↗
PubMed1981

Hydrolysis of guinea pig nascent very low density lipoproteins catalyzed by lipoprotein lipase: activation by hjman apolipoprotein C-II.

Very low density lipoproteins isolated from guinea pig liver perfusate (VLDLp) lack the equivalent of human apolipoprotein C-II (apoC-II), the activator of lipoprotein lipase (LpL). These lipoproteins are therefore ideal substrates with which to investigate the mechanism by which apoC-II activates the enzyme. VLDLp binds apoC-II, and apoC-II associated with VLDLp markedly increases the rate of lipoprotein lipase-catalyzed hydrolysis of VLDLp-triglycerides. The activator potency of apoC-II is independent of the method of enrichment of VLDLp with apoC-II: delipidated human apoC-II and apoC-II transferred from human high density lipoproteins activate lipoprotein lipase to equal extents. ApoC-II causes pH-dependent changes in both apparent Km and VmaX of LpL-catalyzed hydrolysis of VLDLp-triglycerides. At pH l7.4--7.5, the major effects of apoC-II is to decrease the apparent Km by 3.3--4.0 fold. The apparent Vmax is increased 1.3-fold. At pH 6.5 and 8.5, the decrease of apparent Km is less marked, 1.6-fold and 1.4-fold, respectively. At pH 6.5, apoC-II increases the apparent Vmax ty 1.3-fold, while at pH 8.5 the primary effect of apoC-II is a 1.6-fold increase of apparent Vmax. Based on a simple kinetic model, the data suggest that apoC-II favors direct interaction between enzyme and triglyceride within the lipoprotein particle, as well as subsequent catalytic turnover.

Animals↗