Fo membrane domain of ATP synthase from bovine heart mitochondria: purification, subunit composition, and reconstitution with F1-ATPase.


:The Fo membrane domain of the F1Fo-ATP synthase complex has been purified from bovine heart mitochondria. The purification procedure involves the removal of peripheral membrane proteins, including F1-ATPase, from submitochondrial particles with guanidine hydrochloride, followed by extraction of Fo and other membrane proteins from the stripped membranes in the presence of the detergent n-dodecyl beta-D-maltoside. Fo was then purified by ion-exchange and dye ligand chromatography in the presence of the same detergent. Approximately 15 mg of pure Fo was recovered from 1.8 g of mitochondrial membrane protein. The purified Fo is a complex of nine different polypeptides. They are subunits a, b, c, d, e, F6, and A6L characterized before in F1Fo-ATPase preparations, and two new hitherto undetected subunits, named f and g. The sequences of subunits f and g have been determined. They are not related significantly to any known protein, but subunit f appears to contain a membrane-spanning alpha-helix. Proteins f and g are also present in approximately stoichiometric amounts in a highly purified preparation of intact F1Fo-ATPase, and so it is concluded that they are authentic subunits of the bovine enzyme with unknown functions. Dibutyltin 3-hydroxyflavone, an inhibitor of F1Fo-ATPase, also binds to the purified Fo in detergent and competes for binding with venturicidin. In the presence of F1 and OSCP, the purified Fo was reassembled into the intact F1Fo-ATPase complex. Therefore, this procedure provides a relatively abundant source of pure and functional Fo that is suitable for structural analysis.






Collinson IR,Runswick MJ,Buchanan SK,Fearnley IM,Skehel JM,van Raaij MJ,Griffiths DE,Walker JE




Has Abstract


1994-06-28 00:00:00












  • Spin equilibrium in human methemoglobin: effects of inositol hexaphosphate and bezafibrate as measured by resonance Raman spectroscopy.

    abstract::The spin equilibria of several derivatives of human methemoglobin were probed by resonance Raman scattering. The intensity of lines in the Raman spectrum gives a measure of the high-spin (S = 5/2) to low-spin (S = 1/2) ratio which agrees well with the spin equilibria determined from direct magnetic susceptibility meas...


    pub_type: 杂志文章


    authors: Noble RW,DeYoung A,Rousseau DL

    更新日期:1989-06-13 00:00:00

  • Iron regulatory element and internal loop/bulge structure for ferritin mRNA studied by cobalt(III) hexammine binding, molecular modeling, and NMR spectroscopy.

    abstract::The ferritin IRE, a highly conserved (96-99% in vertebrates) mRNA translation regulatory element in animal mRNA, was studied by molecular modeling (using MC-SYM and DOCKING) and by NMR spectroscopy. Cobalt(III) hexammine was used to model hydrated Mg2+. IRE isoforms in other mRNAs regulate mRNA translation or stabilit...


    pub_type: 杂志文章


    authors: Gdaniec Z,Sierzputowska-Gracz H,Theil EC

    更新日期:1998-02-10 00:00:00

  • Purification and characterization of a transmembrane domain-deleted form of lecithin retinol acyltransferase.

    abstract::Lecithin retinol acyltransferase (LRAT) catalyzes the esterification of all-trans-retinol into all-trans-retinyl ester, an essential reaction in the vertebrate visual cycle. Since all-trans-retinyl esters are the substrates for the isomerization reaction that generates 11-cis-retinoids, this esterification reaction is...


    pub_type: 杂志文章


    authors: Bok D,Ruiz A,Yaron O,Jahng WJ,Ray A,Xue L,Rando RR

    更新日期:2003-05-27 00:00:00

  • Structural locations and functional roles of new subsites S5, S6, and S7 in memapsin 2 (beta-secretase).

    abstract::Memapsin 2 (beta-secretase) is the membrane-anchored aspartic protease that initiates the cleavage of beta-amyloid precursor protein (APP), leading to the production of amyloid-beta (Abeta), a major factor in the pathogenesis of Alzheimer's disease. The active site of memapsin 2 has been shown, with kinetic data and c...


    pub_type: 杂志文章


    authors: Turner RT 3rd,Hong L,Koelsch G,Ghosh AK,Tang J

    更新日期:2005-01-11 00:00:00

  • Stopped-flow and steady-state study of the diphenolase activity of mushroom tyrosinase.

    abstract::The reaction of mushroom (Agaricus bisporus) tyrosinase with dioxygen in the presence of several o-diphenolic substrates has been studied by steady-state and transient-phase kinetics in order to elucidate the rate-limiting step and to provide new insights into the mechanism of oxidation of these substrates. A kinetic ...


    pub_type: 杂志文章


    authors: Rodríguez-López JN,Fenoll LG,García-Ruiz PA,Varón R,Tudela J,Thorneley RN,García-Cánovas F

    更新日期:2000-08-29 00:00:00

  • Design of the helix-turn-helix motif: nonlocal effects of quaternary structure in DNA recognition investigated by laser Raman spectroscopy.

    abstract::The operator-binding domain of phage lambda repressor provides a model for DNA recognition by the helix-turn-helix (HTH) motif. In the wild-type protein, dimerization is mediated by hydrophobic packing (of the dyad-related helix 5), which serves as an indirect determinant of operator affinity. The mutant repressor, Ty...


    pub_type: 杂志文章


    authors: Benevides JM,Weiss MA,Thomas GJ Jr

    更新日期:1991-05-07 00:00:00

  • Lung surfactant proteins, SP-B and SP-C, alter the thermodynamic properties of phospholipid membranes: a differential calorimetry study.

    abstract::The ability of the low molecular weight lung surfactant-associated proteins, SP-B and SP-C, to alter the thermotropic properties of synthetic multilamellar vesicles was tested using differential scanning calorimetry (DSC). The presence of either SP-B or SP-C in dipalmitoylphosphatidylcholine (DPPC) or dipalmitoylphosp...


    pub_type: 杂志文章


    authors: Shiffer K,Hawgood S,Haagsman HP,Benson B,Clements JA,Goerke J

    更新日期:1993-01-19 00:00:00

  • Aminophosphonate inhibitors of dialkylglycine decarboxylase: structural basis for slow binding inhibition.

    abstract::The kinetics of inhibition of dialkylglycine decarboxylase by five aminophosphonate inhibitors are presented. Two of these [(R)-1-amino-1-methylpropanephosphonate and (S)-1-aminoethanephosphonate] are slow binding inhibitors. The inhibitors follow a mechanism in which a weak complex is rapidly formed, followed by slow...


    pub_type: 杂志文章


    authors: Liu W,Rogers CJ,Fisher AJ,Toney MD

    更新日期:2002-10-15 00:00:00

  • Electrostatic guidance of glycosyl cation migration along the reaction coordinate of uracil DNA glycosylase.

    abstract::The DNA repair enzyme uracil DNA glycosylase has been crystallized with a cationic 1-aza-2'-deoxyribose-containing DNA that mimics the ultimate transition state of the reaction in which the water nucleophile attacks the anomeric center of the oxacarbenium ion-uracil anion reaction intermediate. Comparison with substra...


    pub_type: 杂志文章


    authors: Bianchet MA,Seiple LA,Jiang YL,Ichikawa Y,Amzel LM,Stivers JT

    更新日期:2003-11-04 00:00:00

  • Ff gene 5 single-stranded DNA-binding protein assembles on nucleotides constrained by a DNA hairpin.

    abstract::The gene 5 protein (g5p) encoded by filamentous Ff phages is an ssDNA-binding protein, which binds to and sequesters the nascent ssDNA phage genome in the process of phage morphogenesis. The g5p also binds with high affinity to DNA and RNA sequences that form G-quadruplex structures. However, sequences that would form...


    pub_type: 杂志文章


    authors: Wen JD,Gray DM

    更新日期:2004-03-09 00:00:00

  • Stoichiometry and specificity of lipid-protein interaction with myelin proteolipid protein studied by spin-label electron spin resonance.

    abstract::The interaction of spin-labeled lipids with the myelin proteolipid apoprotein in complexes with dimyristoylphosphatidylcholine of varying lipid/protein ratios has been studied with electron spin resonance spectroscopy. A first shell of approximately 10 lipids per 25 000-dalton protein is found to be motionally restric...


    pub_type: 杂志文章


    authors: Brophy PJ,Horváth LI,Marsh D

    更新日期:1984-02-28 00:00:00

  • Design, Synthesis, and Potency of Pyruvate Dehydrogenase Complex E1 Inhibitors against Cyanobacteria.

    abstract::Safe and effective algaecides are needed to control agriculturally and environmentally significant algal species. Four series (6, 10, 17, and 21) of 29 novel 4-aminopyrimidine derivatives were rationally designed and synthesized. A part of 10, 17, and 21 displayed potent inhibition of Escherichia coli pyruvate dehydro...


    pub_type: 杂志文章


    authors: Zhou Y,Feng J,He H,Hou L,Jiang W,Xie D,Feng L,Cai M,Peng H

    更新日期:2017-12-12 00:00:00

  • Contribution of the prothrombin fragment 2 domain to the function of factor Va in the prothrombinase complex.

    abstract::The prothrombinase complex assembles through reversible interactions between factor Xa, factor Va and acidic phospholipid-containing membranes in the presence of calcium ions. This complex catalyses the conversion of prothrombin to thrombin through two proteolytic steps. We have used prethrombin 2 as a substrate analo...


    pub_type: 杂志文章


    authors: Krishnaswamy S,Walker RK

    更新日期:1997-03-18 00:00:00

  • Molecular structure and polymorphic map of the human phenylalanine hydroxylase gene.

    abstract::Human phenylalanine hydroxylase is a liver-specific enzyme that catalyzes the conversion of phenylalanine to tyrosine. Absence of enzymatic activity results in phenylketonuria, a genetic disorder that causes development of severe mental retardation in untreated children. In this paper we report the cloning and structu...


    pub_type: 杂志文章


    authors: DiLella AG,Kwok SC,Ledley FD,Marvit J,Woo SL

    更新日期:1986-02-25 00:00:00

  • Kinetic studies of guinea pig liver transglutaminase reveal a general-base-catalyzed deacylation mechanism.

    abstract::Guinea pig liver transglutaminase (TGase) reacts with 0.1 mM N-Cbz-L-Glu(gamma-p-nitrophenyl ester)Gly (5, prepared herein, K(M) = 0.02 mM) to undergo rapid acylation that can be followed spectrophotometrically at 400 nm (pH 7.0, 25 degrees C). Deacylation of the transiently formed thiolester acyl enzyme intermediate ...


    pub_type: 杂志文章


    authors: Leblanc A,Gravel C,Labelle J,Keillor JW

    更新日期:2001-07-27 00:00:00

  • Targeting of protein phosphatases PP2A and PP2B to the C-terminus of the L-type calcium channel Ca v1.2.

    abstract::The L-type Ca(2+) channel Ca(v)1.2 forms macromolecular signaling complexes that comprise the β(2) adrenergic receptor, trimeric G(s) protein, adenylyl cyclase, and cAMP-dependent protein kinase (PKA) for efficient signaling in heart and brain. The protein phosphatases PP2A and PP2B are part of this complex. PP2A coun...


    pub_type: 杂志文章


    authors: Xu H,Ginsburg KS,Hall DD,Zimmermann M,Stein IS,Zhang M,Tandan S,Hill JA,Horne MC,Bers D,Hell JW

    更新日期:2010-12-07 00:00:00

  • Refinement of netropsin bound to DNA: bias and feedback in electron density map interpretation.

    abstract::The X-ray crystal structure of the complex of the B-DNA dodecamer CGCGAATTCGCG with the antitumor drug netropsin has been reexamined to locate the drug accurately for computer-based drug design. The optimum solution is with the drug centered in the AATT region of the minor groove, making three good bifurcated hydrogen...


    pub_type: 杂志文章


    authors: Goodsell DS,Kopka ML,Dickerson RE

    更新日期:1995-04-18 00:00:00

  • Computational study of the resistance shown by the subtype B/HIV-1 protease to currently known inhibitors.

    abstract::Human immunodeficiency virus type 1 protease (HIV-1 PR) is an essential enzyme in the HIV-1 life cycle. As such, this protein represents a major drug target in AIDS therapy, but emerging resistance to antiretroviral inhibitor cocktails, caused by high viral mutation rates, represents a significant challenge in AIDS tr...


    pub_type: 杂志文章


    authors: Genoni A,Morra G,Merz KM Jr,Colombo G

    更新日期:2010-05-18 00:00:00

  • Mechanism of tubulin assembly: guanosine 5'-triphosphate hydrolysis decreases the rate of microtubule depolymerization.

    abstract::The rate of depolymerization of microtubules upon lowering the temperature was found to depend on the amount of time elapsed since the beginning of the assembly process. In the first minutes following self-assembly at 37 degrees C, microtubules are more cold sensitive and depolymerize faster than later at the steady s...


    pub_type: 杂志文章


    authors: Bonne D,Pantaloni D

    更新日期:1982-03-02 00:00:00

  • Increased β-Sheet Dynamics and D-E Loop Repositioning Are Necessary for Cu(II)-Induced Amyloid Formation by β-2-Microglobulin.

    abstract::β-2-Microglobulin (β2m) forms amyloid fibrils in the joints of patients undergoing dialysis treatment as a result of kidney failure. One of the ways in which β2m can be induced to form amyloid fibrils in vitro is via incubation with stoichiometric amounts of Cu(II). To better understand the structural changes caused b...


    pub_type: 杂志文章


    authors: Borotto NB,Zhang Z,Dong J,Burant B,Vachet RW

    更新日期:2017-02-28 00:00:00

  • Mechanistic studies on full length and the catalytic domain of the tandem SH2 domain-containing protein tyrosine phosphatase: analysis of phosphoenzyme levels and Vmax stimulatory effects of glycerol and of a phosphotyrosyl peptide ligand.

    abstract::SHP-1, a protein tyrosine phosphatase containing two tandem SH2 domains, is autoinhibited at rest by its N-terminal SH2 (N-SH2) domain. Relief from autoinhibition and a subsequent 10-60-fold increase in V(max) have been observed upon N-SH2 domain engagement by a specific phosphotyrosyl ligand or upon deletion of the S...


    pub_type: 杂志文章


    authors: Wang J,Walsh CT

    更新日期:1997-03-11 00:00:00

  • Crystal structure of rat apo-heme oxygenase-1 (HO-1): mechanism of heme binding in HO-1 inferred from structural comparison of the apo and heme complex forms.

    abstract::Heme oxygenase (HO) catalyzes the oxidative cleavage of heme to biliverdin by utilizing O(2) and NADPH. HO (apoHO) was crystallized as twinned P3(2) with three molecules per asymmetric unit, and its crystal structure was determined at 2.55 A resolution. Structural comparison of apoHO and its complex with heme (HO-heme...


    pub_type: 杂志文章


    authors: Sugishima M,Sakamoto H,Kakuta Y,Omata Y,Hayashi S,Noguchi M,Fukuyama K

    更新日期:2002-06-11 00:00:00

  • Properties of Intermediates in the Catalytic Cycle of Oxalate Oxidoreductase and Its Suicide Inactivation by Pyruvate.

    abstract::Oxalate:ferredoxin oxidoreductase (OOR) is an unusual member of the thiamine pyrophosphate (TPP)-dependent 2-oxoacid:ferredoxin oxidoreductase (OFOR) family in that it catalyzes the coenzyme A (CoA)-independent conversion of oxalate into 2 equivalents of carbon dioxide. This reaction is surprising because binding of C...


    pub_type: 杂志文章


    authors: Pierce E,Mansoorabadi SO,Can M,Reed GH,Ragsdale SW

    更新日期:2017-06-06 00:00:00

  • Evidence that the principal CoII-binding site in human serum albumin is not at the N-terminus: implication on the albumin cobalt binding test for detecting myocardial ischemia.

    abstract::Human serum albumin (HSA) is the most abundant protein in the blood plasma and is involved in the transport of metal ions. Four metal-binding sites with different specificities have been described in HSA: (i) the N-terminal site provided by Asp1, Ala2, and His3, (ii) the site at the reduced Cys34, (iii) site A, includ...


    pub_type: 杂志文章


    authors: Mothes E,Faller P

    更新日期:2007-02-27 00:00:00

  • Role of metal ions in catalysis by enolase: an ordered kinetic mechanism for a single substrate enzyme.

    abstract::Spectroscopic and kinetic methods have been used to explore the roles of divalent metal ions in the enolase-catalyzed dehydration of 2-phosphoglycerate (2-PGA). Enolase requires 2 equiv of metal ion per active site for maximal activity. Previous crystallographic studies [Larsen, T. M., Wedekind, J. E., Rayment, I., an...


    pub_type: 杂志文章


    authors: Poyner RR,Cleland WW,Reed GH

    更新日期:2001-07-10 00:00:00

  • Evidence for coupled motion and hydrogen tunneling of the reaction catalyzed by glutamate mutase.

    abstract::Glutamate mutase is one of a group of adenosylcobalamin-dependent enzymes that catalyze unusual isomerizations that proceed through organic radical intermediates generated by homolytic fission of the coenzyme's unique cobalt-carbon bond. These enzymes are part of a larger family of enzymes that catalyze radical chemis...


    pub_type: 杂志文章


    authors: Cheng MC,Marsh EN

    更新日期:2007-01-23 00:00:00

  • Carbon monoxide binding to human hemoglobin A0.

    abstract::The carbon monoxide binding curve to human hemoglobin A0 has been measured to high precision in experimental conditions of 600 microM heme, 0.1 M N-(2-hydroxyethyl)piperazine-N'-2-ethanesulfonic acid, 0.1 M NaCl, 10 mM inositol hexaphosphate, 1 mM disodium ethylenediaminetetraacetic acid, pH 6.94, and 25 degrees C. Co...


    pub_type: 杂志文章


    authors: Di Cera E,Doyle ML,Connelly PR,Gill SJ

    更新日期:1987-10-06 00:00:00

  • 3 Beta-hydroxy-delta 5-steroid dehydrogenase/3-keto-delta 5-steroid isomerase from bovine adrenals: mechanism of inhibition by 3-oxo-4-aza steroids and kinetic mechanism of the dehydrogenase.

    abstract::Several 3-oxo-4-aza steroids (1) have been identified as inhibitors of the 3 beta-hydroxy-delta 5-steroid dehydrogenase/3-keto-delta 5-steroid isomerase catalyzed conversion of pregnenolone to progesterone. By kinetically decoupling the two enzyme activities isolated from bovine adrenal cortex, it has been demonstrate...


    pub_type: 杂志文章


    authors: Brandt M,Levy MA

    更新日期:1989-01-10 00:00:00

  • Catalysis of phosphoryl group transfer. The role of divalent metal ions in the hydrolysis of lactic acid O-phenyl phosphate and salicylic acid O-aryl phosphates.

    abstract::The spontaneous hydrolyses of lactic acid O-phenyl phosphate (I) and, to a lesser extent, 3-hydroxybutyric acid O-phenyl phosphate (II) have been investigated and compared with similar intramolecular and bimolecular reactions. Compared to bimolecular nucleophilic reactions, the reactivity of II is similar to other sys...


    pub_type: 杂志文章


    authors: Steffens JJ,Siewers IJ,Benkovic SJ

    更新日期:1975-06-03 00:00:00

  • The crystal structure of human procathepsin K.

    abstract::Cathepsin K is a cysteine protease present in human osteoclasts that plays an important role in bone resorption. Cathepsin K is synthesized as an inactive proenzyme and activated under conditions of low pH. Autoproteolytic processing of the N-terminal 99 amino acid propeptide produces the active, mature form of cathep...


    pub_type: 杂志文章


    authors: LaLonde JM,Zhao B,Janson CA,D'Alessio KJ,McQueney MS,Orsini MJ,Debouck CM,Smith WW

    更新日期:1999-01-19 00:00:00