scholarly journals Acyl-CoA binding and acylation of UDP-glucuronosyltransferase isoforms of rat liver: their effect on enzyme activity

1995 ◽  
Vol 312 (1) ◽  
pp. 301-308 ◽  
Author(s):  
A Yamashita ◽  
M Watanabe ◽  
T Tonegawa ◽  
T Sugiura ◽  
K Waku
Keyword(s):  
Fatty Acid ◽  
Rat Liver ◽  
Liver Microsomes ◽  
Fatty Acyl ◽  
Binding Activity ◽  
Rat Liver Microsomes ◽  
Sequencing Analysis ◽  
Terminal Amino ◽  
Udp Glucuronosyltransferase ◽  
Hydrolysis Of

When [14C]arachidonoyl-CoA was incubated with crude extracts of rat liver microsomes, [14C]arachidonic acid was incorporated into many proteins, suggesting that modification of these proteins with fatty acid, i.e. acylation, occurred. Using a [14C]arachidonyl-CoA labelling assay, 50 and 53 kDa proteins were purified from rat liver microsomes to near homogeneity by sequential chromatography on Red-Toyopearl, hydroxyapatite, heparin-Toyopearl, Blue-Toyopearl and UDP-hexanolamine-agarose. Acylation of the 50 and 53 kDa proteins occurred in the absence of any other protein, suggesting that these molecules catalyse autoacylation. The acylation was dependent on the length of the incubation period and the concentration of [14C]arachidonoyl-CoA. The 50 and 53 kDa proteins also had acyl-CoA-binding activity; initial rates of acyl-CoA binding and acylation were 0.25 and 0.004 min-1 respectively. The proteins also had weak but distinct acyl-CoA-hydrolysing activity (0.006 min-1). These results suggest that the proteins catalysed the sequential reactions of binding to acyl-CoA, autoacylation, and hydrolysis of fatty acid. N-terminal amino acid sequencing analysis showed these proteins to be UDP-glucuronosyltransferase (UDPGT) isoforms. UDPGT activity was inhibited by arachidonoyl-CoA. These results suggest that binding of acyl-CoA and acylation of UDPGT isoforms regulate the enzyme activities, implying a possible novel function for fatty acyl-CoA in glucuronidation, which is involved in the metabolism of drugs, steroids and bilirubin.

scholarly journals Fatty acyl-CoA esters inhibit glucose-6-phosphatase in rat liver microsomes

1995 ◽  
Vol 307 (2) ◽  
pp. 391-397 ◽  
Author(s):  
R Fulceri ◽  
A Gamberucci ◽  
H M Scott ◽  
R Giunti ◽  
A Burchell ◽  
...  
Keyword(s):  
Palmitic Acid ◽  
Phosphatase Activity ◽  
Rat Liver ◽  
Liver Microsomes ◽  
Fatty Acyl ◽  
Rat Liver Microsomes ◽  
Control Value ◽  
Scattering Measurements ◽  
Induced Changes ◽  
Enzymic Assay

In native rat liver microsomes glucose 6-phosphatase activity is dependent not only on the activity of the glucose-6-phosphatase enzyme (which is lumenal) but also on the transport of glucose-6-phosphate, phosphate and glucose through the respective translocases T1, T2 and T3. By using enzymic assay techniques, palmitoyl-CoA or CoA was found to inhibit glucose-6-phosphatase activity in intact microsomes. The effect of CoA required ATP and fatty acids to form fatty acyl esters. Increasing concentrations (2-50 microM) of CoA (plus ATP and 20 microM added palmitic acid) or of palmitoyl-CoA progressively decreased glucose-6-phosphatase activity to 50% of the control value. The inhibition lowered the Vmax without significantly changing the Km. A non-hydrolysable analogue of palmitoyl-CoA also inhibited, demonstrating that binding of palmitoyl-CoA rather than hydrolysis produces the inhibition. Light-scattering measurements of osmotically induced changes in the size of rat liver microsomal vesicles pre-equilibrated in a low-osmolality buffer demonstrated that palmitoyl-CoA alone or CoA plus ATP and palmitic acid altered the microsomal permeability to glucose 6-phosphate, but not to glucose or phosphate, indicating that T1 is the site of palmitoyl-CoA binding and inhibition of glucose-6-phosphatase activity in native microsomes. The type of inhibition found suggests that liver microsomes may comprise vesicles heterogeneous with respect to glucose-6-phosphate translocase(s), i.e. sensitive or insensitive to fatty acid ester inhibition.

Inhibitory effect of curcumin on fatty acid desaturation inMortierella alpina 1S-4 and rat liver microsomes

Lipids ◽  
1992 ◽  
Vol 27 (7) ◽  
pp. 509-512 ◽  
Author(s):  
Sakayu Shimizu ◽  
Saeree Jareonkitmongkol ◽  
Hiroshi Kawashima ◽  
Kengo Akimoto ◽  
Hideaki Yamada
Keyword(s):  
Fatty Acid ◽  
Rat Liver ◽  
Liver Microsomes ◽  
Inhibitory Effect ◽  
Fatty Acid Desaturation ◽  
Rat Liver Microsomes

Fluorescence polarization studies of rat liver microsomes with altered phospholipid desaturase activities

1980 ◽  
Vol 58 (10) ◽  
pp. 952-958 ◽  
Author(s):  
E. L. Pugh ◽  
M. Kates ◽  
A. G. Szabo
Keyword(s):  
Fatty Acid ◽  
Rat Liver ◽  
Fluorescence Polarization ◽  
Desaturase Activity ◽  
Liver Microsomes ◽  
Rat Liver Microsomes ◽  
Polarization Ratio ◽  
Parinaric Acid ◽  
Cis And Trans ◽  
Normal Controls

Phospholipid desaturase activity of rat liver microsomes can be varied by dietary manipulation: rats starved and refed a "fat-free" diet have two to three times the activity of normal controls whereas starved rats have no detectable activity. These changes were accompanied by changes in fatty acid composition of the microsomal phospholipids, resulting in a double bond:saturated fatty acid mole ratio (moles double bonds per mole saturated fatty acid) of 5.7 in control and starved rats and 4.7 in starved–refed rats. Fluorescence polarization ratio [Formula: see text] of cis- and trans-parinaric acid (PnA) showed no significant differences in physical state of the three microsomal preparations. However, the isolated microsomal phospholipids with trans-PnA as probe showed differences in the temperature at which onset of a change in polarization ratio occurred (starved–refed > normal > starved rats). With the cis-PnA as probe, the polarization ratio showed no change in the range 10–40 °C but was significantly higher (1.8) in starved–refed rats than in normal and starved rats (1.6 in both cases). These data indicate that the microsomal phospholipids of starved–refed animals were in a less fluid state than those from control and starved rats and that this decrease in fluidity was correlated with an increase in phospholipid desaturase activity.

A Study on the Effect of Lysolecithin and Phospholipase A on Membrane-Bound Galactosyltransferase

1974 ◽  
Vol 52 (11) ◽  
pp. 1053-1066 ◽  
Author(s):  
Sailen Mookerjea ◽  
James W. M. Yung
Keyword(s):  
Liver Microsomes ◽  
Fatty Acyl ◽  
Phospholipase A ◽  
Synthetase Activity ◽  
Rat Liver Microsomes ◽  
Physiological Concentration ◽  
Activation Effect ◽  
Membrane Bound ◽  
Hydrolysis Of

Addition of lysolecithin caused very marked activation of UDP-galactose:glycoprotein galactosyltransferase in rat liver microsomes and in Golgi-rich membranes. Lysolecithin activated galactosyltransferase when the enzyme was assayed both with endogenous acceptor and with exogenous proteins or monosaccharides as acceptors. Lactose synthetase activity in presence of α-lactalbumin was also stimulated by lysolecithin. Lecithin, lysophosphatidylethanolamine, lysophosphatidic acid, and glycerophosphorylcholine did not activate the enzyme, suggesting that both fatty acyl and phosphorylcholine groups of the lysolecithin molecule are required for the observed activation. The degree of activation was about the same when myristoyl-, palmitoyl-, oleoyl-, or stearoyllysolecithin were tested. The activation by lysolecithin was observed well within the physiological concentration of the lipid in the liver cell. Saturating amounts of Triton masked the effect of lysolecithin.Brief preincubation with phospholipase A activated the enzyme and generated lysolecithin in the membranes. Triton and lysolecithin activated the enzyme without any lag time, whereas phospholipase A activation was dependent on preincubation and also on an alkaline pH favorable for the hydrolysis of phospholipid. EDTA blocked the activation effect of phospholipase A but had no effect on activation by lysolecithin. Albumin and cholesterol opposed the effects of lysolecithin and phospholipase A on the enzyme. Two successive incubations of the microsomes with lysolecithin caused considerable release of the enzyme into the soluble fraction. The role of lysolecithin in the activation of the enzyme is probably related to the solubilization of the membrane and consequent enhanced interaction of the enzyme with substrate. Lysolecithin also activated N-acetylglucosaminyl- and sialyltransferase activities in microsomes. A possible role of lysolecithin is indicated in the regulation of glycosylation reactions in mammalian system.

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