A Substrate- and Position-specific Acylation of sn-Glycerol 3-Phosphate by Rat Liver Mitochondria

Monroy, Gladys; Rola, F.H.; Pullman, Maynard E.

doi:10.1016/s0021-9258(19)44668-1

Cited by 132 publications

(15 citation statements)

References 61 publications

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“…The substrate specificity determined with our preparation is similar to that reported with crude rat liver mitochondrial extracts. Overall, mitochondrial GPAT prefers saturated fatty acyl-CoA as a substrate (Monroy et al, 1973;Kelker & Pullman, 1979;Monroy et al, 1972). The preference for saturated fatty acyl donors was also demonstrated in E. coli GPAT, consistent with the known positional distribution of saturated fatty acids (Green et al, 1981).…”

Section: Purification and Reconstitution Of Gpat Expressed In Sf9supporting

confidence: 73%

Purification and Reconstitution of Murine Mitochondrial Glycerol-3-phosphate Acyltransferase. Functional Expression in Baculovirus-Infected Insect Cells

Yet¹,

Moon²,

Sul³

1995

Biochemistry

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Glycerol-3-phosphate acyltransferase (GPAT) catalyzes the initial step in glycerolipid biosynthesis. We recently cloned a cDNA to a 6.8-kb mRNA, a message that can be induced dramatically by feeding a high-carbohydrate diet [Paulauskis & Sul (1988) J. Biol. Chem. 263, 7049-7054; Shin et al. (1991) J. Biol. Chem. 266, 23834-23839], and identified the open reading frame, p90, as mitochondrial GPAT [Yet et al. (1993) Biochemistry 32, 9486-9491]. To initiate characterization of mitochondrial GPAT, we purified and reconstituted the GPAT activity using phospholipids after expressing functional enzyme in Sf9 insect cells. Infection with recombinant virus containing p90 sequence resulted in high levels of GPAT expression in mitochondria, compared to noninfected cells or cells infected with the reverse orientation insertion baculovirus. There was a dramatic increase in N-ethylmaleimide-resistant mitochondrial GPAT activity. The GPAT protein was not detectable by Western blot in noninfected Sf9 cells or in cells infected with the GPAT sequence in the reverse orientation. However, in cells infected with GPAT in the correct orientation, there was a dramatic increase in the GPAT protein that was readily detectable by Coomassie staining both in total extracts and in the mitochondrial fraction. To ease the purification, we next expressed GPAT as a polyhistidine fusion protein in insect cells. The polyhistidine tag did not interfere with targeting to mitochondria or with the catalytic activity of GPAT.(ABSTRACT TRUNCATED AT 250 WORDS)

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Section: Purification and Reconstitution Of Gpat Expressed In Sf9supporting

confidence: 73%

Purification and Reconstitution of Murine Mitochondrial Glycerol-3-phosphate Acyltransferase. Functional Expression in Baculovirus-Infected Insect Cells

Yet¹,

Moon²,

Sul³

1995

Biochemistry

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“…Mice deficient in acyl-CoA:diacylglycerol transferase 1 (DGAT1), cat-duced fasting concentrations of diacylglycerol, further implicating this metabolite as a potentially important mediator of fat-alyzing the final step in the glycerol 3-phosphate pathway, share several phenotypic similarities with mtGPAT1 −/− mice induced hepatic insulin resistance. These data also suggest that mtGPAT1 might be a potent therapeutic target to treat he- (Chen et al, 2002;Smith et al, 2000). DGAT1 −/− mice are lean, have smaller fat pads, and lack alterations in plasma glu-patic steatosis and hepatic insulin resistance.…”

Section: Resultsmentioning

confidence: 95%

“…cose, insulin, and fatty-acid concentrations. But in contrast to mechanism involving increased energy expenditure and activity mice were singly housed, and in order to minimize stress to the animals, experiments were carried out in several separate but identically treated (Chen et al, 2002;Smith et al, 2000). DGAT2, the second chargroups of mice.…”

Section: Resultsmentioning

confidence: 99%

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Prevention of hepatic steatosis and hepatic insulin resistance in mitochondrial acyl-CoA:glycerol-sn-3-phosphate acyltransferase 1 knockout mice

et al. 2005

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In order to investigate the role of mitochondrial acyl-CoA:glycerol-sn-3-phosphate acyltransferase 1 (mtGPAT1) in the pathogenesis of hepatic steatosis and hepatic insulin resistance, we examined whole-body insulin action in awake mtGPAT1 knockout (mtGPAT1(-/-)) and wild-type (wt) mice after regular control diet or three weeks of high-fat feeding. In contrast to high-fat-fed wt mice, mtGPAT1(-/-) mice displayed markedly lower hepatic triacylglycerol and diacylglycerol concentrations and were protected from hepatic insulin resistance possibly due to a lower diacylglycerol-mediated PKC activation. Hepatic acyl-CoA has previously been implicated in the pathogenesis of insulin resistance. Surprisingly, compared to wt mice, mtGPAT1(-/-) mice exhibited increased hepatic insulin sensitivity despite an almost 2-fold elevation in hepatic acyl-CoA content. These data suggest that mtGPAT1 might serve as a novel target for treatment of hepatic steatosis and hepatic insulin resistance and that long chain acyl-CoA's do not mediate fat-induced hepatic insulin resistance in this model.

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“…Eicosapentaenoic acid, DPAn-3, and DHA all increased in the TG pool following supplemental FO (Figure 4); however, the increase was modest relative to the increase observed for the PL pool (Figure 2). Polyunsaturated FA, particularly >20 carbon PUFA, were preferentially incorporated into the sn-2 position of PL (Monroy et al, 1972); therefore, the potential to alter the >20 carbon PUFA composition of the PL fraction is greater. In hepatocytes, the absolute increase in DHA was greater than for EPA, which contrasted with both plasma and peripheral blood mononuclear cell pools in calves supplemented with a similar FO (M. A.…”

Section: Discussionmentioning

confidence: 99%

Effects of dietary supplemental fish oil during the peripartum period on blood metabolites and hepatic fatty acid compositions and total triacylglycerol concentrations of multiparous Holstein cows

Ballou¹,

Gomes

Juchem

et al. 2009

Journal of Dairy Science

102

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The objectives were to evaluate the effects of dietary fish oil on plasma metabolite, hepatic fatty acid composition, and total triacylglycerol concentrations. Multiparous Holstein cows (n = 42) were completely randomized to 1 of 3 treatments at 3 wk prepartum. Treatments were no supplemental lipid or supplemental lipid from either Energy Booster (Milk Specialties Co., Dundee, IL) or fish oil. Treatment diets were fed from -21 d relative to expected date of parturition until 10 d postpartum. Treatments were fed as a bolus before the a.m. feeding. The dose of lipid fed during the prepartum period was 250 g, whereas approximately 0.92% of the previous day's dry matter intake was supplemented postpartum. Blood was collected 3 times weekly for determination of plasma metabolites. Liver biopsies were performed at 21 and 10 d before expected date of parturition and 1 and 14 d after parturition to determine fatty acid compositions and total triacylglycerol concentrations. Dry matter intake, milk yield, and loss of body weight or body condition score were not affected by supplementing the diet with lipid or by the source of lipid. Supplemental lipid tended to increase plasma glucose and decrease nonesterified fatty acids during the postpartum period. Furthermore, plasma beta-hydroxybutyrate was reduced during the postpartum period in the lipid-supplemented treatments. However, source of supplemental lipid had no influence on any blood metabolite. Supplemental fish oil altered the fatty acid composition of liver phospholipids and triacylglycerols, decreasing total saturated fatty acids and increasing total n-3 and long-chain polyunsaturated fatty acids (>20 carbon fatty acids). Despite the altered fatty acid composition, hepatic total triacylglycerol concentrations were unaffected by supplemental fish oil. Furthermore, the improved metabolic profile following lipid supplementation did not decrease hepatic total triacylglycerol concentrations.

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A Substrate- and Position-specific Acylation of sn-Glycerol 3-Phosphate by Rat Liver Mitochondria

Cited by 132 publications

References 61 publications

Purification and Reconstitution of Murine Mitochondrial Glycerol-3-phosphate Acyltransferase. Functional Expression in Baculovirus-Infected Insect Cells

Purification and Reconstitution of Murine Mitochondrial Glycerol-3-phosphate Acyltransferase. Functional Expression in Baculovirus-Infected Insect Cells

Prevention of hepatic steatosis and hepatic insulin resistance in mitochondrial acyl-CoA:glycerol-sn-3-phosphate acyltransferase 1 knockout mice

Effects of dietary supplemental fish oil during the peripartum period on blood metabolites and hepatic fatty acid compositions and total triacylglycerol concentrations of multiparous Holstein cows

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