Phospholipid asymmetry in large unilamellar vesicles induced by transmembrane pH gradients.


:The influence of membrane pH gradients on the transbilayer distribution of some common phospholipids has been investigated. We demonstrate that the transbilayer equilibrium of the acidic phospholipids egg phosphatidylglycerol (EPG) and egg phosphatidic acid (EPA) can be manipulated by membrane proton gradients, whereas phosphatidylethanolamine, a zwitterionic phospholipid, remains equally distributed between the inner and outer monolayers of large unilamellar vesicles (LUVs). Asymmetry of EPG is examined in detail and demonstrated by employing three independent techniques: ion-exchange chromatography, 13C NMR, and periodic acid oxidation of the (exterior) EPG headgroup. In the absence of a transmembrane pH gradient (delta pH) EPG is equally distributed between the outer and inner monolayers of LUVs. When vesicles composed of either egg phosphatidylcholine (EPC) or DOPC together with 5 mol % EPG are prepared with a transmembrane delta pH (inside basic, outside acidic), EPG equilibrates across the bilayer until 80-90% of the EPG is located in the inner monolayer. Reversing the pH gradient (inside acidic, outside basic) results in the opposite asymmetry. The rate at which EPG equilibrates across the membrane is temperature dependent. These observations are consistent with a mechanism in which the protonated (neutral) species of EPG is able to traverse the bilayer. Under these circumstances EPG would be expected to equilibrate across the bilayer in a manner that reflects the transmembrane proton gradient. A similar mechanism has been demonstrated to apply to simple lipids that exhibit weak acid or base characteristics [Hope, M. J., & Cullis, P. R. (1987) J. Biol. Chem 262, 4360-4366]






Hope MJ,Redelmeier TE,Wong KF,Rodrigueza W,Cullis PR




Has Abstract


1989-05-16 00:00:00












  • Time-resolved rotational dynamics of phosphorescent-labeled myosin heads in contracting muscle fibers.

    abstract::We have measured the microsecond rotational motions of myosin heads in contracting rabbit psoas muscle fibers by detecting the transient phosphorescence anisotropy of eosin-5-maleimide attached specifically to the myosin head. Experiments were performed on small bundles (10-20 fibers) of glycerinated rabbit psoas musc...


    pub_type: 杂志文章


    authors: Stein RA,Ludescher RD,Dahlberg PS,Fajer PG,Bennett RL,Thomas DD

    更新日期:1990-10-30 00:00:00

  • Selectivity of fluorescent lipid analogues for lipid domains.

    abstract::We have examined the phase partition preferences of the even chain length (n = 10-22) diacyl-3'3'-indocarbo-cyanine iodides (Cn diI) incorporated in disaturated lecithin (PC) vesicles. Two parameters were used to determine this phase preference: (i) the direction of shift of the phase transition temperature (Tm) induc...


    pub_type: 杂志文章


    authors: Klausner RD,Wolf DE

    更新日期:1980-12-23 00:00:00

  • Barbiturate inhibition of GLUT-1 mediated hexose transport in human erythrocytes exhibits substrate dependence for equilibrium exchange but not unidirectional sugar flux.

    abstract::Barbiturates inhibit GLUT-1 mediated hexose transport both in vivo [Gjedde & Rasmussen (1980) J. Neurochem. 35, 1382-1387; Otsuka et al. (1991) Am. J. Physiol. 261, R265-R275] and in vitro [Honkanen et al. (1995) Biochemistry 34, 535-544]. In the present study, the mechanism by which barbiturates inhibit GLUT-1 mediat...


    pub_type: 杂志文章


    authors: el-Barbary A,Fenstermacher JD,Haspel HC

    更新日期:1996-12-03 00:00:00

  • Membrane trafficking of aquaporin 1 is mediated by protein kinase C via microtubules and regulated by tonicity.

    abstract::It is well-known that the rapid flow of water into and out of cells is controlled by membrane proteins called aquaporins (AQPs). However, the mechanisms that allow cells to quickly respond to a changing osmotic environment are less well established. Using GFP-AQP fusion proteins expressed in HEK293 cells, we demonstra...


    pub_type: 杂志文章


    authors: Conner MT,Conner AC,Brown JE,Bill RM

    更新日期:2010-02-09 00:00:00

  • A hexameric helicase encircles one DNA strand and excludes the other during DNA unwinding.

    abstract::The bacteriophage T7 DNA helicase/primase (gene 4 protein) is a ring-like hexamer that encircles ssDNA and requires forked DNA to catalyze DNA unwinding. We report that optimal rates of unwinding of forked DNA require ssDNA tails of 55 nucleotides on the 5'-to-3' strand and 15 nucleotides on the 3'-to-5' strand. Surpr...


    pub_type: 杂志文章


    authors: Hacker KJ,Johnson KA

    更新日期:1997-11-18 00:00:00

  • Crystal structure of murine 11 beta-hydroxysteroid dehydrogenase 1: an important therapeutic target for diabetes.

    abstract::11Beta-hydroxysteroid dehydrogenase type 1 (11beta-HSD1) catalyzes the conversion of 11-dehydrocorticosterone to its active form corticosterone in rodents (or cortisone to cortisol in humans). The reductive reaction of the 11-keto to 11-hydroxyl is the pivotal switch in the activation of glucocorticoids. An excess of ...


    pub_type: 杂志文章


    authors: Zhang J,Osslund TD,Plant MH,Clogston CL,Nybo RE,Xiong F,Delaney JM,Jordan SR

    更新日期:2005-05-10 00:00:00

  • How Do Branched Detergents Stabilize GPCRs in Micelles?

    abstract::The structural and functional properties of G protein-coupled receptors (GPCRs) are often studied in a detergent micellar environment, but many GPCRs tend to denature or aggregate in short alkyl chain detergents. In our previous work [Lee, S., et al. (2016) J. Am. Chem. Soc.138, 15425-15433], we showed that GPCRs in a...


    pub_type: 杂志文章


    authors: Lee S,Ghosh S,Jana S,Robertson N,Tate CG,Vaidehi N

    更新日期:2020-06-16 00:00:00

  • Complex structures between the N-type calcium channel (CaV2.2) and ω-conotoxin GVIA predicted via molecular dynamics.

    abstract::The N-type voltage-gated Ca(2+) channel CaV2.2 is one of the important targets for pain management. ω-Conotoxins isolated from venoms of cone snails, which specifically inhibit CaV2.2, are promising scaffolds for novel analgesics. The inhibitory action of ω-conotoxins on CaV2.2 has been examined experimentally, but th...


    pub_type: 杂志文章


    authors: Chen R,Chung SH

    更新日期:2013-05-28 00:00:00

  • Nuclear Overhauser experiments at 500 MHz on the downfield proton spectra of 5S ribonucleic acid and its complex with ribosomal protein L25.

    abstract::The downfield (9-15 ppm) proton spectrum of Escherichia coli 5S RNA has been examined at 500 MHz by using nuclear Overhauser methods. The data confirm the existence of the terminal and procaryotic loop helices within the molecule [Fox, G. E., & Woese, C. R. (1975) Nature (London) 256, 505-506]. Very little stable, dou...


    pub_type: 杂志文章


    authors: Kime MJ,Moore PB

    更新日期:1983-05-24 00:00:00

  • Mode of interaction of aminooxy compounds with sheep liver serine hydroxymethyltransferase.

    abstract::The interaction of aminooxy compounds such as aminooxyacetate (AAA), L-canaline, and hydroxylamine with sheep liver serine hydroxymethyltransferase (EC was studied by absorption spectra and stopped-flow spectrophotometry and compared with the unique feature of interaction of O-amino-D-serine (OADS) with the e...


    pub_type: 杂志文章


    authors: Baskaran N,Prakash V,Savithri HS,Radhakrishnan AN,Appaji Rao N

    更新日期:1989-12-12 00:00:00

  • Structural investigation of the binding of a herpesviral protein to the SH3 domain of tyrosine kinase Lck.

    abstract::Herpesvirus saimiri codes for a tyrosine kinase interacting protein (Tip) that interacts with both the SH3 domain and the kinase domain of the T-cell-specific tyrosine kinase Lck via two separate motifs. The activation of Lck by Tip is considered as a key event in the transformation of human T-lymphocytes during herpe...


    pub_type: 杂志文章


    authors: Schweimer K,Hoffmann S,Bauer F,Friedrich U,Kardinal C,Feller SM,Biesinger B,Sticht H

    更新日期:2002-04-23 00:00:00

  • Sulfide dehydrogenase activity of the monomeric flavoprotein SoxF of Paracoccus pantotrophus.

    abstract::Flavocytochrome c-sulfide dehydrogenases (FCSDs) are complexes of a flavoprotein with a c-type cytochrome performing hydrogen sulfide-dependent cytochrome c reduction in vitro. The amino acid sequence analysis revealed that the phylogenetic relationship of different flavoproteins reflected the relationship of sulfur-o...


    pub_type: 杂志文章


    authors: Quentmeier A,Hellwig P,Bardischewsky F,Wichmann R,Friedrich CG

    更新日期:2004-11-23 00:00:00

  • Conformational characteristics of luliberin. Luminescence properties at liquid-nitrogen temperature.

    abstract::The luminescence properties (fluorescence and phosphorescence) of luliberin have been investigated at liquid-nitrogen temperature (77 K) in 50% ethylene glycolaqueous buffer at various pH's. Calculation of the energy-transfer efficiency between Trp and Tyr residues leads to an evaluation of the distance separating the...


    pub_type: 杂志文章


    authors: Marche P,Montenay-Garestier T,Fromageot P,Hélène C

    更新日期:1976-12-28 00:00:00

  • Molten globule formation in apomyoglobin monitored by the fluorescent probe Nile Red.

    abstract::The interaction of nile red (NR) with apomyoglobin (ApoMb) in the native (pH 7) and molten globule (pH 4) states was investigated using experimental and computational methods. NR binds to hydrophobic locations in ApoMb with higher affinity (K(d) = 25 +/- 5 microM) in the native state than in the molten globule state (...


    pub_type: 杂志文章


    authors: Polverini E,Cugini G,Annoni F,Abbruzzetti S,Viappiani C,Gensch T

    更新日期:2006-04-25 00:00:00

  • Kinetics of the subtransition of asymmetrically substituted phosphatidylcholines.

    abstract::The thermodynamics and kinetics of the subtransition L epsilon----P beta of sonicated unilamellar vesicles of 1-myristoyl-2-stearoylphosphatidylcholine (1M-2S-PC) and of 1-stearoyl-2-myristoylphosphatidylcholine (1S-2M-PC) were studied by equilibrium cooling curves and temperature-jump relaxation spectrometry with an ...


    pub_type: 杂志文章


    authors: Tümmler B,Herrmann U,Maass G,Eibl H

    更新日期:1984-08-28 00:00:00

  • Fluorescent and photoactivatable fluorescent derivatives of tetrodotoxin to probe the sodium channel of excitable membranes.

    abstract::Fluorescent and photoactivatable fluorescent derivatives of tetrodotoxin (TTX) have been synthesized. N-Methylanthraniloylglycine hydrazide, anthraniloyl hydrazide, and 2-azidoanthraniloylglycine hydrazide were coupled to the carbonyl at C6 of oxidized tetrodotoxin to form stable fluorescent hydrazones. The C6 ketone ...


    pub_type: 杂志文章


    authors: Angelides KJ

    更新日期:1981-07-07 00:00:00

  • Direct spectroscopic and kinetic evidence for the involvement of a peroxodiferric intermediate during the ferroxidase reaction in fast ferritin mineralization.

    abstract::Rapid freeze-quench (RFQ) Mössbauer and stopped-flow absorption spectroscopy were used to monitor the ferritin ferroxidase reaction using recombinant (apo) frog M ferritin; the initial transient ferric species could be trapped by the RFQ method using low iron loading (36 Fe2+/ferritin molecule). Biphasic kinetics of f...


    pub_type: 杂志文章


    authors: Pereira AS,Small W,Krebs C,Tavares P,Edmondson DE,Theil EC,Huynh BH

    更新日期:1998-07-14 00:00:00

  • DNA translocation by human uracil DNA glycosylase: the case of single-stranded DNA and clustered uracils.

    abstract::Human uracil DNA glycosylase (hUNG) plays a central role in DNA repair and programmed mutagenesis of Ig genes, requiring it to act on sparsely or densely spaced uracil bases located in a variety of contexts, including U/A and U/G base pairs, and potentially uracils within single-stranded DNA (ssDNA). An interesting qu...


    pub_type: 杂志文章


    authors: Schonhoft JD,Stivers JT

    更新日期:2013-04-16 00:00:00

  • Determination of proton transfer rates by chemical rescue: application to bacterial reaction centers.

    abstract::The bacterial reaction center (RC) converts light into chemical energy through the reduction of an internal quinone molecule Q(B) to Q(B)H(2). In the native RC, proton transfer is coupled to electron transfer and is not rate-controlling. Consequently, proton transfer is not directly observable, and its rate was unknow...


    pub_type: 杂志文章


    authors: Paddock ML,Adelroth P,Feher G,Okamura MY,Beatty JT

    更新日期:2002-12-17 00:00:00

  • An amino acid residue (S201) in the retinal binding pocket regulates the photoreaction pathway of phoborhodopsin.

    abstract::Phoborhodopsin from Halobacterium salinarum (salinarum phoborhodopsin, spR also called HsSR II) is a photoreceptor for the negative phototaxis of the bacterium. A unique feature of spR is the formation of a shorter wavelength photoproduct, P480, observed at liquid nitrogen temperature beside the K intermediate. Format...


    pub_type: 杂志文章


    authors: Dai G,Zhang Y,Tamogami J,Demura M,Kamo N,Kandori H,Iwasa T

    更新日期:2011-08-23 00:00:00

  • Transfer RNA-pseudouridine synthetase Pus1 of Saccharomyces cerevisiae contains one atom of zinc essential for its native conformation and tRNA recognition.

    abstract::RNA:pseudouridine synthetase (Pus1) from Saccharomyces cerevisiae is a multisite specific enzyme that catalyzes the formation of pseudouridine at positions 34 and 36 of intron-containing precursor tRNAIle and at positions 27 and/or 28 of several yeast tRNAs. In this paper we demonstrate that the purified recombinant P...


    pub_type: 杂志文章


    authors: Arluison V,Hountondji C,Robert B,Grosjean H

    更新日期:1998-05-19 00:00:00

  • Isolation and characterization of bovine factor XII (Hageman factor).

    abstract::Factor XII was purified approximately 14 000-fold from bovine plasma by ammonium sulfate fractionation followed by heparin-agarose, DEAE-Sephadex, CM-cellulose, arginine-agarose, and benzamidine-agarose column chromatography. By this method, about 15 mg of protein was purified from 15 L of plasma with an overall yield...


    pub_type: 杂志文章


    authors: Fujikawa K,Walsh KA,Davie EW

    更新日期:1977-05-17 00:00:00

  • Mechanism of action of thrombin on fibrinogen. Kinetic evidence for involvement of aspartic acid at position P10.

    abstract::The following peptide was synthesized by classical methods in solution: Ac-Asp-Phe-Leu-Ala-Glu-Gly-Gly-Gly-Val-Arg-Gly-Pro-Arg-Val-NHCH3 (F-8). The Michaelis-Menten parameters for the hydrolysis of the Arg-Gly bond in F-8 by thrombin were determined to be Kcat = 31 X 10(-11) M [(NIH unit/L) s]-1 and KM = 310 X 10(-6) ...


    pub_type: 杂志文章


    authors: Marsh HC Jr,Meinwald YC,Thannhauser TW,Scheraga HA

    更新日期:1983-08-30 00:00:00

  • Studies of early events of folding of a predominately β-sheet protein using fluorescence correlation spectroscopy and other biophysical methods.

    abstract::The interplay between the early collapse of the unfolded state and the formation of the secondary structure has been the subject of extensive research in protein chemistry. In this study, we used the intestinal fatty acid binding protein (IFABP), a small model protein with predominately β-sheet structure, to study the...


    pub_type: 杂志文章


    authors: Sarkar S,Chattopadhyay K

    更新日期:2014-03-11 00:00:00

  • Biotin synthase is a pyridoxal phosphate-dependent cysteine desulfurase.

    abstract::Biotin synthase (BioB) is an iron-sulfur dimeric enzyme which catalyzes the last step in biotin synthesis. The reaction consists of the introduction of a sulfur atom into dethiobiotin. It is shown here that BioB displays a significant cysteine desulfurase activity, providing it with the ability to mobilize sulfur from...


    pub_type: 杂志文章


    authors: Ollagnier-De-Choudens S,Mulliez E,Hewitson KS,Fontecave M

    更新日期:2002-07-23 00:00:00

  • The Cys-Tyr cross-link of cysteine dioxygenase changes the optimal pH of the reaction without a structural change.

    abstract::Cysteine dioxygenase (CDO) is a non-heme monoiron enzyme with an unusual posttranslational modification in the proximity of the ferrous iron active site. This modification, a cysteine to tyrosine thioether bond, cross-links two β-strands of the β-barrel. We have investigated its role in catalysis through a combined cr...


    pub_type: 杂志文章


    authors: Davies CG,Fellner M,Tchesnokov EP,Wilbanks SM,Jameson GN

    更新日期:2014-12-23 00:00:00

  • Structures of apolipoprotein A-II and a lipid-surrogate complex provide insights into apolipoprotein-lipid interactions.

    abstract::Apolipoproteins A-I and A-II form the major protein constituents of high-density lipid particles (HDL), the concentration of which is inversely correlated with the frequency of heart disease in humans. Although the physiological role of apolipoprotein A-II is unclear, evidence for its involvement in free fatty acid me...


    pub_type: 杂志文章,收录出版


    authors: Kumar MS,Carson M,Hussain MM,Murthy HM

    更新日期:2002-10-01 00:00:00

  • Chloroperoxidase, a janus enzyme.

    abstract::Chloroperoxidase is a versatile fungal heme-thiolate protein that catalyzes a variety of one-electron and two-electron oxidations. We report here that the alkylation of an essential histidine residue showed no effect on the one-electron peroxidations but inhibited two-electron oxidations. The pH profiles of different ...


    pub_type: 杂志文章


    authors: Manoj KM,Hager LP

    更新日期:2008-03-04 00:00:00

  • Purification and partial characterization of a DNA 3'-phosphatase from Schizosaccharomyces pombe.

    abstract::Cells that depend on oxygen for survival constantly produce reactive oxygen species that attack DNA to produce a variety of lesions, including single-strand breaks with 3'-blocking groups such as 3'-phosphate and 3'-phosphoglycolate. These 3'-blocking ends prevent the activity of DNA polymerase and are generally remov...


    pub_type: 杂志文章


    authors: Jilani A,Ramotar D

    更新日期:2002-06-18 00:00:00

  • Effects of oxidation on the structure and stability of human low-density lipoprotein.

    abstract::Oxidation of low-density lipoprotein (LDL), the major cholesterol carrier in plasma, is thought to promote atherogenesis via several mechanisms. One proposed mechanism involves fusion of oxidized LDL in the arterial wall; another involves oxidation-induced amyloid formation by LDL apolipoprotein B. To test these mecha...


    pub_type: 杂志文章


    authors: Jayaraman S,Gantz DL,Gursky O

    更新日期:2007-05-15 00:00:00