E J McMurchie

The Commonwealth Scientific and Industrial Research Organisation, Canberra, Australian Capital Territory, Australia

Are you E J McMurchie?

Claim your profile

Publications (93)227.94 Total impact

  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The protein CrV2 is encoded by a polydnavirus integrated into the genome of the endoparasitoid Cotesia rubecula (Hymenoptera:Braconidae:Microgastrinae) and is expressed in host larvae with other gene products of the polydnavirus to allow successful development of the parasitoid. CrV2 expression has previously been associated with immune suppression, although the molecular basis for this was not known. Here, we have used time-resolved Förster resonance energy transfer (TR-FRET) to demonstrate high affinity binding of CrV2 to Gα subunits (but not the Gβγ dimer) of heterotrimeric G-proteins. Signals up to 5-fold above background were generated, and an apparent dissociation constant of 6.2 nm was calculated. Protease treatment abolished the TR-FRET signal, and the presence of unlabeled CrV2 or Gα proteins also reduced the TR-FRET signal. The activation state of the Gα subunit was altered with aluminum fluoride, and this decreased the affinity of the interaction with CrV2. It was also demonstrated that CrV2 preferentially bound to Drosophila Gα(o) compared with rat Gα(i1). In addition, three CrV2 homologs were detected in sequences derived from polydnaviruses from Cotesia plutellae and Cotesia congregata (including the immune-related early expressed transcript, EP2). These data suggest a potential mode-of-action of immune suppressors not previously reported, which in addition to furthering our understanding of insect immunity may have practical benefits such as facilitating development of novel controls for pest insect species.
    Journal of Biological Chemistry 01/2011; 286(12):10466-75. · 4.65 Impact Factor
  • Source
    Tamara Cooper, Edward J McMurchie, Wayne R Leifert
    [Show abstract] [Hide abstract]
    ABSTRACT: The [(35)S]GTPgammaS binding assay to measure G protein activation following agonist binding to G protein-coupled receptors (GPCRs) remains a powerful molecular technique to substantiate traditional pharmacological values of potency, efficacy, and affinity. The method described uses membrane preparations of the alpha(2A)-adrenergic receptor and purified G protein subunits expressed in Sf9 cells, reconstituted into a functional signaling system. This technology is generic and could be used with other GPCRs to demonstrate initial signaling events following receptor activation. Agonist-stimulated [(35)S]GTPgammaS binding is measured in a 96-well plate format using scintillation counting.
    Methods in molecular biology (Clifton, N.J.) 02/2009; 552:143-51. · 1.29 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Expression of proteins in insect cells using recombinant baculoviruses has gained wide use in the G protein-coupled receptor (GPCR) community. This expression system produces high yields of functional receptor, is able to perform post-translational modifications, and is readily adaptable to large-scale culture. Here, we describe the generic methods for expressing a GPCR using baculovirus-infected insect cells, including the maintenance of insect cell culture. Data are presented for polyhedrin promoter-driven expression of a C-terminal 6 x histidine-tagged mammalian M(2) muscarinic receptor in Sf9 cells. Results demonstrate that expressed receptor could be detected and quantified using radiolabeled ligand binding, that expression was maximal at approximately 72 h post-infection, and that expression levels could be altered by addition of various ligands to cultures of infected insect cells.
    Methods in molecular biology (Clifton, N.J.) 02/2009; 552:115-29. · 1.29 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Lanthanide-binding tags (LBTs) are small peptides showing great promise for use in novel imaging technologies due to their high affinity for terbium, europium and gadolinium ions which have unique luminescent and/or paramagnetic properties. When fused to functional proteins the LBT can act as a site directed location for the binding of lanthanide ions which could be utilised in assays (usually luminescence-based) to derive data about protein interactions and functionality. This study shows the lanthanide-binding tag, LBT2, to have an apparent Kd for terbium of 9.3 nM. Other common ions showed much lower affinities for LBT2; however, gadolinium provided competitive binding with an IC50 of 13.7 nM. LBTs were then fused to both the M2-muscarinic receptor and a chimeric Gα-subunit allowing them to be labelled with terbium. The presence of this LBT did not disrupt either the ligand binding or signalling of the receptor. The construction of a LBT motif in association with specific proteins presents exciting possibilities to employ bimodal (luminescent and magnetic) imaging properties in relation to such areas as biosensor development and design.
    Journal of Bionanoscience 05/2008; 2(1):27-34.
  • [Show abstract] [Hide abstract]
    ABSTRACT: Attachment of histidine-tagged proteins to nitrilotriacetic acid (NTA) surfaces via a chelated nickel(II) ion is a powerful technique for immobilizing proteins in an oriented fashion. In this study, mixed monolayers of NTA disulfide containing triethylene glycol spacers were self-assembled on gold substrates, and nickel(II) ions were chelated to bind Gα-proteins containing either 6- or 16-residue N-terminal histidine tags. Both proteins were shown to be functional, in terms of their ability to bind BODIPYFL-GTPγS. Surface plasmon resonance was used to show that the monolayer containing NTA disulfide displays a stronger binding surface for histidine-tagged Gαi1 protein compared with monovalent NTA attached to a dextran matrix, as the 6-histidine Gαi1 was unable to be stably bound to the NTA-dextran surface under the required buffer conditions. The 16-residue histidine tag displayed a stronger binding to the mixed monolayer compared with the 6-residue histidine tag, which is the standard length tag commonly used for controlled nickel-histidine interactions. This stronger binding demonstrates that mixed self-assembled monolayers of bivalent NTA and triethylene glycol display an enhanced tethering to proteins that contain additional histidine residues. This may facilitate the assembly of functional biosensors, in that directed attachment of active proteins to surfaces (in formats such as plates, beads, and microarrays) may be conducted in a more robust fashion.
    Journal of Bionanoscience 05/2007; 1(1):22-30.
  • Source
    Edward McMurchie, Wayne Leifert
    [Show abstract] [Hide abstract]
    ABSTRACT: Signal transduction by G-protein coupled receptors (GPCRs) underpins a multitude of physiological processes. Ligand recognition by the receptor leads to the activation of a generic molecular switch involving heterotrimeric G-proteins and guanine nucleotides. With growing interest and commercial investment in GPCRs in areas such as drug targets, orphan receptors, high-throughput screening of drugs and biosensors, greater attention will focus on assay development to allow for miniaturization, ultrahigh-throughput and, eventually, microarray/biochip assay formats that will require nanotechnology-based approaches. Stable, robust, cell-free signaling assemblies comprising receptor and appropriate molecular switching components will form the basis of future GPCR/G-protein platforms, which should be able to be adapted to such applications as microarrays and biosensors. This chapter focuses on cell-free GPCR assay nanotechnologies and describes some molecular biological approaches for the construction of more sophisticated, surface-immobilized, homogeneous, functional GPCR sensors. The latter points should greatly extend the range of applications to which technologies based on GPCRs could be applied.
    Springer Handbook of Nanotechnology, ISBN 978-3-540-29855-7. Springer-Verlag Berlin Heidelberg, 2007, p. 505. 01/2007;
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The ability to express and purify modified recombinant proteins, so they retain their biological function in a cell-free format, has provided a basis for development of molecular biosensors. Here we utilize recombinant G protein-coupled receptors (GPCRs) and their G proteins for cell-free detection of various binding partners. Fusion peptides were used to improve surface-attachment and fluorescent-labelling capabilities. A novel homogeneous fluorescence resonance energy transfer (FRET)-based assay was developed to detect rearrangements in the G protein heterotrimer. By using this heterotrimeric 'molecular switch', we are developing a generic technology such that multiple GPCRs could be assayed for ligand-mediated activation while tethered to surfaces or in solution, with increased throughput compared to current assay platforms.
    Australian Journal of Chemistry 01/2007; 60:309-313. · 1.87 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: Functionalised gold nanoparticles, capable of specifically binding histidine-tagged proteins, are of interest as their unique optical properties can facilitate the study of protein-protein interactions. Citrate-stabilized gold nanoparticles were functionalised with surface capping agents to specifically bind histidine-tagged proteins, and to test the stability of the nanoparticle solutions under various conditions.
    Nanoscience and Nanotechnology, 2006. ICONN '06. International Conference on; 08/2006
  • [Show abstract] [Hide abstract]
    ABSTRACT: The ability to express and purify modified recombinant signalling proteins such that they retain their biological function in a cell-free context has provided a basis for production of molecular biosensors. Here the authors utilise G-protein coupled receptors (GPCRs) and their G-proteins to detect various binding partners in a cell-free environment. Molecular biology approaches were employed to express these proteins using baculovirus and bacteria, and to alter their characteristics to improve surface-attachment and fluorescent labelling capabilities. Ligand-mediated signalling of a GPCR could be measured (using [<sup>35</sup>S]GTPgammaS-binding assays) in a reconstituted system with recombinant proteins either free in solution or attached to Ni<sup>2+</sup>-coated beads. Affinity of histidine-tagged proteins for a Ni<sup>2+</sup>-coated surface was significantly enhanced by addition of extra histidine residues to the tag, as determined by surface plasmon resonance. This was due to the longer tag occupying, on average, a greater number of available histidine-binding sites. Further, a novel homogeneous fluorescence resonance energy transfer (FRET)-based assay has been developed to detect rearrangements in the G-protein heterotrimer. Investigation of small peptides that can be fused to G-protein subunits, allowing for site-specific fluorescent labelling, was undertaken in order to improve the resolution of the "first generation" FRET assay. By utilizing this improved G-protein heterotrimer "molecular switch", we are developing a generic technology such that a range of GPCRs could be assayed for ligand-mediated activation while attached to surfaces (e.g. on beads or as microarrays) or in solution (e.g. multi-well plates), with increased throughput.
    Nanoscience and Nanotechnology, 2006. ICONN '06. International Conference on; 08/2006
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: G-protein-coupled receptors transduce their signals through G-protein subunits which in turn are subject to modulation by other intracellular proteins such as the regulators of G-protein signaling (RGS) proteins. We have developed a cell-free, homogeneous (mix and read format), time-resolved fluorescence resonance energy transfer (TR-FRET) assay to monitor heterotrimeric G-protein subunit interactions and the interaction of the G alpha subunit with RGS4. The assay uses a FRET pair consisting of a terbium cryptate chelate donor spectrally matched to an Alexa546 fluor acceptor, each of which is conjugated to separate protein binding partners, these being G alpha(i1):beta4gamma2 or G alpha(i1):RGS4. Under conditions favoring specific binding between labeled partners, high-affinity interactions were observed as a rapid increase (>fivefold) in the FRET signal. The specificity of these interactions was demonstrated using denaturing or competitive conditions which caused significant reductions in fluorescence (50-85%) indicating that labeled proteins were no longer in close proximity. We also report differential binding effects as a result of altered activation state of the G alpha(i1) protein. This assay confirms that interactions between G-protein subunits and RGS4 can be measured using TR-FRET in a cell- and receptor-free environment.
    Analytical Biochemistry 08/2006; 355(2):201-12. · 2.58 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The ability to express and purify modified recombinant signalling proteins such that they retain their biological function in a cell-free context, has provided a basis for production of molecular biosensors. Here we utilise G-protein Coupled Receptors (GPCRs) and their G-proteins to detect various binding partners in a cell-free environment. Molecular biology approaches were employed to express these proteins using baculovirus and bacteria, and to alter their characteristics to improve surface-attachment and fluorescent labelling capabilities. Ligand-mediated signalling of a GPCR could be measured (using [ 35 S]GTPγS-binding assays) in a reconstituted system with recombinant proteins either free in solution or attached to Ni 2+ -coated beads. Affinity of histidine-tagged proteins for a Ni 2+ -coated surface was significantly enhanced by addition of extra histidine residues to the tag, as determined by surface plasmon resonance. This was due to the longer tag occupying, on average, a greater number of available histidine-binding sites. Further, a novel homogeneous fluorescence resonance energy transfer (FRET)-based assay has been developed to detect rearrangements in the G-protein heterotrimer. Investigation of small peptides that can be fused to G-protein subunits, allowing for site-specific fluorescent labelling, was undertaken in order to improve the resolution of the "first generation" FRET assay. By utilizing this improved G-protein heterotrimer "molecular switch", we are developing a generic technology such that a range of GPCRs could be assayed for ligand-mediated activation while attached to surfaces (e.g. on beads or as microarrays) or in solution (e.g. multi-well plates), with increased throughput.
    International Conference on Nanoscience and Nanotechnology, Brisbane, Australia; 07/2006
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The n-3 polyunsaturated fatty acids (PUFAs) found in fish oil (FO) have been shown to protect against reperfusion arrhythmias, a manifestation of reperfusion injury, which is believed to be induced by the formation of reactive oxygen species (ROS) and intracellular calcium (Ca2+) overload. Adult rats fed a diet supplemented with 10% FO had a higher proportion of myocardial n-3 PUFAs and increased expression of antioxidant enzymes compared with the saturated fat (SF)-supplemented group. Addition of hydrogen peroxide (H2O2) to cardiomyocytes isolated from rats in the SF-supplemented group increased the proportions of cardiomyocytes contracting in an asynchronous manner, increased the rate of Ca2+ influx, and increased the diastolic and systolic [Ca2+]i compared with the FO group. H2O2 exposure increased the membrane fluidity of cardiomyocytes from the FO group. These results demonstrate that dietary FO supplementation is associated with a reduction in the susceptibility of myocytes to ROS-induced injury and this may be related to membrane incorporation of n-3 PUFAs, increased antioxidant defenses, changes in cardiomyocyte membrane fluidity, and the ability to prevent rises in cellular Ca2+ in response to ROS.
    Free Radical Biology and Medicine 06/2006; 40(9):1592-602. · 5.27 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Signal transduction by G-protein-coupled receptors (GPCRs) underpins a multitude of physiological processes. Ligand recognition by the receptor leads to activation of a generic molecular switch involving heterotrimeric G-proteins and guanine nucleotides. Signal transduction has been studied extensively with both cell-based systems and assays comprising isolated signaling components. Interest and commercial investment in GPCRs in areas such as drug targets, orphan receptors, high throughput screening, biosensors, and so on will focus greater attention on assay development to allow for miniaturization, ultra-high throughput and, eventually, microarray/biochip assay formats. Although cell-based assays are adequate for many GPCRs, it is likely that these formats will limit the development of higher density GPCR assay platforms mandatory for other applications. Stable, robust, cell-free signaling assemblies comprising receptor and appropriate molecular switching components will form the basis of future GPCR assay platforms adaptable for such applications as microarrays. The authors review current cell-free GPCR assay technologies and molecular biological approaches for construction of novel, functional GPCR assays.
    Journal of Biomolecular Screening 01/2006; 10(8):765-79. · 2.21 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: G-protein coupled receptors (GPCRs) form a ternary complex of agonist, receptor and G-proteins during primary signal transduction at the cell membrane. Downstream signalling is thought to be preceded by the process of dissociation of Galpha and Gbetagamma subunits, thus exposing new surfaces to interact with downstream effectors. We demonstrate here for the first time, the dissociation of heterotrimeric G-protein subunits (i.e., Galpha and Gbetagamma) following agonist-induced GPCR (alpha(2A)-adrenergic receptor; alpha(2A)-AR) activation in a cell-free assay system. alpha(2A)-AR membranes were reconstituted with the G-proteins (+/-hexahistidine-tagged) Galpha(i1) and Gbeta1gamma2 and functional signalling was determined following activation of the reconstituted receptor:G-protein complex with the potent agonist UK-14304, and [35S]GTPgammaS. In the presence of Ni(2+)-coated agarose beads, the activated his-tagged Galpha(i1)his-[35S]GTPgammaS complex was captured on the Ni(2+)-presenting surface. When his-tagged Gbeta1gamma2 (Gbeta1gamma2his) was used with Galpha(i1), the [35S]GTPgammaS-bound Galpha(i1) was not present on the Ni(2+)-coated beads, but rather, it was separated from the beta1gamma2(his)-beads, demonstrating receptor-induced dissociation of Galpha and Gbetagamma subunits. Treatment of the reconstituted alpha(2A)-AR membranes containing Gbeta1gamma2his:Galpha(i1) with imidazole confirmed the specificity for the Ni2+:G-protein surface dissociation of Galpha(i1) from Gbeta1gamma2his. These data demonstrate for the first time, the complete dissociation of the G-protein subunits and extend observations on the role of G-proteins in the assembly and disassembly of the ternary complex in the primary events of GPCR signalling.
    Molecular Membrane Biology 01/2005; 22(6):507-17. · 3.13 Impact Factor
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: The long-chain (n-3) polyunsaturated fatty acids (PUFA) have been reported to exhibit health benefits and healing properties for the gastrointestinal tract. The aim of this study was to investigate the effects of dietary fish oil supplementation on the in vitro contractility of gut tissue. Rats (9 wk old) were fed synthetic diets supplemented with 170 g/kg Sunola oil (SO; 850 g/kg as oleic acid [18:1(n-9)]) or with 100 g/kg of the SO replaced by saturated animal fat (SF) or fish oil (FO) for 4 wk. In the colon, there was no difference in the sensitivity (50% effective concentration) or the maximal contraction among the three dietary groups induced by acetylcholine or 8-iso-prostaglandin (PG)E(2) with the rat colon being relatively insensitive to the thromboxane mimetic U-46619. However, in the ileum, the FO group had greater maximal contractions induced by acetylcholine and 8-iso-PGE(2) compared with the SO and SF groups (P < 0.05), and greater maximal contractions induced by PGE(2), PGF(2alpha) and U-46619 compared with the SF group (P < 0.05). FO feeding increased the incorporation of (n-3) PUFA (eicosapentaenoic [20:5(n-3)], docosapentaenoic [22:5(n-3)] and docosahexaenoic acids [22:6(n-3) primarily at the expense of (n-6) PUFA (linoleic [18:2(n-6)] and arachidonic acids [20:4(n-6)]) in the ileum and colon phospholipid fatty acids (P < 0.05). The FO group had a lower cecal digesta pH (P < 0.001) and a greater butyrate concentration than the SF group (P < 0.05). These results suggest that dietary (n-3) PUFA may modulate the contractility of the small intestine.
    Journal of Nutrition 10/2002; 132(9):2506-13. · 4.20 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: This study examined the effects of dietary incorporation of n-3 polyunsaturated fatty acids (PUFAs) into cardiac membrane phospholipids on Ca2+ handling (using Fura-2) and arrhythmic contractility in electrically-stimulated, adult rat ventricular cardiomyocytes. Dietary lipid supplementation with fish oil (FO) for 3 weeks significantly increased the proportion of total n-3 polyunsaturated fatty acids (in particular, docosahexaenoic acid) in ventricular membrane phospholipids compared with a saturated fat (SF) supplemented diet (26.2 ± 0.9% vs 6.9 ± 0.9%, respectively, P < 0.001). Cardiomyocytes isolated from the FO group were significantly (P < 0.001) less susceptible to isoproterenol-induced arrhythmic contractile activity compared with the SF group over a range of isoproterenol concentrations. Isoproterenol (0.5 μM) stimulation increased end-diastolic and systolic [Ca2+]i to a similar extent in both groups. The time constant of Ca2+ transient decay was significantly increased in the FO group compared with the SF group (98.4 ± 2.8 ms, n = 8 and 86.9 ± 2.1 ms, n = 8, P < 0.01, respectively). The effect of dietary n-3 PUFA incorporation into membrane phospholipids was not associated with changes in sarcoplasmic reticulum Ca2+ content (measured by rapid application of caffeine) or membrane fluidity. The increase in the time constant of decay of Ca2+ transients following dietary supplementation with FO may indicate altered functioning of the sarcolemmal Na+-Ca2+ exchanger by n-3 PUFA incorporation into membrane phospholipids.
    The Journal of nutritional biochemistry 07/2001; · 4.29 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: A modified apparatus is described that provides for the simultaneous bathing of the serosa of an intact piece of isolated guinea pig ileum while allowing infusion of the isolated lumen. The comparative compartmental potency of the opioid agonists morphine, casomorphins, and enkephalins to inhibit electrically driven contractions are described in this system. The rank-order potency for serosally applied opioid agonists was (IC(50) values, nM): [D-Ala(2),N-Me-Phe(4),Gly-ol(5)]-enkephalin (DAMGO) (15)>[D-Ala(2),D-Leu(5)]-enkephalin (DADLE) (35)> or =morphine (46)> or =[D-Ala(2)]-met-enkephalinamide (55)>[D-Ala(2)]-beta-casomorphin[1--4] amide (122)>beta-casomorphin[1--4] amide (940)>met- and leu-enkephalin (>6000). This contrasted to the rank-order potency for the luminally applied opioid agonists: DADLE (63)>DAMGO (135)>[D-Ala(2)]-met-enkephalinamide=morphine (4700)>[D-Ala(2)]-beta-casomorphin[1--4] amide (29000). beta-Casomorphin[1--4] amide, leu-enkephalin and met-enkephalin are mostly inactive when applied luminally. Furthermore, the opioid antagonists, casoxin 4 and [D-Ala(2)]-casoxin 4, when infused into the lumen, significantly overcame the inhibitory effect of morphine added to the serosal side. This model provides an assay and screening system to differentiate between the effects of chemical agents applied via the blood stream (serosa) or food side (lumen) on quiescent or electrically driven gut activity of the nervous plexi or receptor systems of the ileum.
    Journal of Pharmacological and Toxicological Methods 01/2001; 45(1):39-46. · 2.15 Impact Factor
  • [Show abstract] [Hide abstract]
    ABSTRACT: A modified apparatus is described that provides for the simultaneous bathing of the serosa of an intact piece of isolated guinea pig ileum while allowing infusion of the isolated lumen. The comparative compartmental potency of the opioid agonists morphine, casomorphins, and enkephalins to inhibit electrically driven contractions are described in this system. The rank-order potency for serosally applied opioid agonists was (IC50 values, nM): [d-Ala2,N-Me-Phe4,Gly-ol5]-enkephalin (DAMGO) (15)>[d-Ala2,d-Leu5]-enkephalin (DADLE) (35)≥morphine (46)≥[d-Ala2]-met-enkephalinamide (55)>[d-Ala2]-β-casomorphin[1–4] amide (122)>β-casomorphin[1–4] amide (940)>met- and leu-enkephalin (>6000). This contrasted to the rank-order potency for the luminally applied opioid agonists: DADLE (63)>DAMGO (135)>[d-Ala2]-met-enkephalinamide=morphine (4700)>[d-Ala2]-β-casomorphin[1–4] amide (29000). β-Casomorphin[1–4] amide, leu-enkephalin and met-enkephalin are mostly inactive when applied luminally. Furthermore, the opioid antagonists, casoxin 4 and [d-Ala2]-casoxin 4, when infused into the lumen, significantly overcame the inhibitory effect of morphine added to the serosal side. This model provides an assay and screening system to differentiate between the effects of chemical agents applied via the blood stream (serosa) or food side (lumen) on quiescent or electrically driven gut activity of the nervous plexi or receptor systems of the ileum.
    Journal of Pharmacological and Toxicological Methods. 01/2001;
  • Journal of Molecular and Cellular Cardiology - J MOL CELL CARDIOL. 01/2001; 33(6).
  • [Show abstract] [Hide abstract]
    ABSTRACT: A protective effect of the n-3 polyunsaturated fatty acids (PUFAs) in preventing ventricular fibrillation in experimental animals and cultured cardiomyocytes has been demonstrated in a number of studies. In this study, a possible role for the n-3 PUFAs in the treatment of atrial fibrillation (AF) was investigated at the cellular level using atrial myocytes isolated from young adult rats as the experimental model. Electrically-stimulated, synchronously-contracting myocytes were induced to contract asynchronously by the addition of 10 M isoproterenol. Asynchronous contractile activity was reduced following acute addition of the n-3 PUFAs docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) at 10 M, compared with no fatty acid addition (from 99.0 : 1.0% to 30.7 5.2% (p < 0.05)="" for="" dha="" and="" 23.8="" ="" 2.8%="" (p="">< 0.01)="" for="" epa),="" while="" the="" saturated="" fatty="" acid,="" docosanoic="" acid="" (da)="" and="" the="" methyl="" ester="" of="" dha="" (dha="" m.e.)="" did="" not="" exert="" a="" significant="" effect="" on="" asynchronous="" contractile="" activity.="" asynchronous="" contractile="" activity="" was="" also="" reduced="" to="" 1.7="" ="" 1.7%="" in="" the="" presence="" of="" the="" membrane="" fluidising="" agent,="" benzyl="" alcohol="" (p="">< 0.001="" vs="" no="" fatty="" acid="" addition).="" cell="" membrane="" fluidity="" was="" determined="" by="" steady="" state="" fluorescence="" anisotropy="" using="" the="" fluorescent="" probe,="" tmap-dph.="" addition="" of="" dha,="" epa="" or="" benzyl="" alcohol="" significantly="" increased="" sarcolemmal="" membrane="" fluidity="" (decreased="" anisotropy,="">ss) of atrial myocytes compared with no addition of fatty acid (control) (from rss = 0.203 0.004 to 0.159 0.004 (p < 0.01)="" for="" dha,="" 0.166="" ="" 0.001="" (p="">< 0.01)="" for="" epa="" and="" 0.186="" 0.003="" (p="">< 0.05)="" for="" benzyl="" alcohol,="" while="" da="" and="" dha="" m.e.="" were="" without="" effect.="" it="" is="" concluded="" that="" the="" n-3="" pufas="" exert="" anti-asynchronous="" effects="" in="" rat="" atrial="" myocytes="" by="" a="" mechanism="" which="" may="" involve="" changes="" in="" membrane="">
    Molecular and Cellular Biochemistry 01/2000; 206(1):33-41. · 2.33 Impact Factor

Publication Stats

883 Citations
227.94 Total Impact Points

Institutions

  • 2000–2006
    • The Commonwealth Scientific and Industrial Research Organisation
      Canberra, Australian Capital Territory, Australia
  • 1999–2006
    • University of Adelaide
      • Department of Physiology
      Adelaide, South Australia, Australia
  • 1988–1992
    • Flinders Medical Centre
      Tarndarnya, South Australia, Australia
  • 1989
    • University of New England (Australia)
      • Department of Zoology
      Armidale, New South Wales, Australia
  • 1985
    • Flinders University
      • School of Biological Sciences
      Tarndarnya, South Australia, Australia
  • 1984
    • University of Melbourne
      Melbourne, Victoria, Australia
  • 1973–1982
    • Macquarie University
      Sydney, New South Wales, Australia
  • 1981
    • University of Alberta
      Edmonton, Alberta, Canada