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INACTIVATION AND THE SITE Of LABELING Of f1-ATPASE fROM MYCOBACTERIUM-PHLEI BY DICYCLOHEXYLCARBODIIMIDE, 7-CHLORO-4-NITROBENZO-2-OXA-1,3-DIAZOLE AND QUINACRINE MUSTARD

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第1期764卷 105-113页

作者： AGARWAL, N KALRA, VK UNIV SO CALIF SCH MED DEPT BIOCHEM LOS ANGELES CA 90033 USA

The f1-ATPase from M. phlei is inactivated by dicyclohexylcarbodiimide (DCCD), 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) and quinacrine mustard. The inactivation is time- and concentration-dependent and in the case of DCCD being more pronounced at acidic pH. The minimum inactivation half-time (t1/2) for DCCD, NBD-Cl and quinacrine mustard was 14, 6 and 7 min, respectively. Inactivation of f1-ATPase resulted in the incorporation of [14C]DCCD and [14C]NBD-Cl into .alpha. and .gamma. subunits. The incorporation of label into .alpha. and .gamma. subunits, utilizing [14C]NBD-Cl, was reversible by dithiothreitol. Complete inactivation, by linear extrapolation to zero activity, revealed that 4 mol [14C]DCCD and 4 mol [14C]NBD-Cl bind/mol f1-ATPase. Kinetic and binding studies show that these probes bind to site(s) distinct from ATP-binding site in f1-ATPase from M. phlei.

关键词： Dicyclohexylcarbodiimide Quinacrine mustard Enzyme inactivation f 1 -ATPase (M. phlei) DCCD dicyclohexylcarbodiimide EDAC 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide EEDQ N -(ethoxycarbonyl)-2-ethoxy-1,2-dihydroquinoline Mes 4-morpholineethanesulphonic acid NBD-Cl 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole TEMED N , N , N ′, N ′-tetramethylethylenediamine

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ISOLATION Of THE MITOCHONDRIAL f1-f0 ADENOSINE-TRIPHOSPHATASE BY SEPHAROSE-HEXYLAMMONIUM CHROMATOGRAPHY - PROPERTIES AND RECONSTITUTION IN LIPOSOMES

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ANALYTICAL BIOCHEMISTRY 1984年第1期142卷 215-220页

作者： DREYfUS, G CELIS, H RAMIREZ, J Departamento de Bioenergética Centro de Investigaciones en Fisiología Celular Universidad Nacional Autonoma de Mexico Apartado Postal 70-600 04510 México D.F. Mexico

Lauryl dimethylamino oxide, a zwitterionic detergent, was employed to solubilize the H+ ATPase from beef heart mitochondria. A simple preparation procedure was devised to obtain f1-f0 based on a method described to purify f1 ATPase which consists of the selective adsorption of f1 to Sepharose-hexylammonium beads. The preparation showed .apprx. 18 bands in sodium dodecyl sulfate-polyacrylamide gel electrophoresis;5 correspond to f1 subunits and the rest probably to the stalk and hydrophobic sector f0. The binding of [14C]dicyclohexylcarbodiimide to a low-molecular-weight component of this preparation was demonstrated. The f1-f0 complex was reconstituted into phospholipid vesicles which displayed ATP-Pi exchange and ATP-dependent 9-aminoacridine fluorescence quenching, both sensitive to proton channel inhibitors.

关键词： f 1 -f 0 ATPase reconstitution affinity chromatography mitochondria membrane proteins

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TWO-DIMENSIONAL GEL-ELECTROPHORESIS Of MEMBRANE-PROTEINS USING ANIONIC AND CATIONIC DETERGENTS - APPLICATION TO THE STUDY Of MITOCHONDRIAL f0-f1-ATPASE

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第2期775卷 239-245页

作者： PENIN, f GODINOT, C GAUTHERON, DC Laboratoire de Biologie et Technologie des Membranes du CNRS Université Claude Bernard de Lyon F 69622 Villeurbanne Cedex France

Polyacrylamide gel electrophoresis in the presence of a cationic detergent, tetradecyltrimethylammonium bromide (TDAB) has been compared to electrophoresis in the presence of an anionic detergent, sodium dodecyl sulfate (SDS). Although, in both systems, the peptides generally migrated as a function of their molecular weight, the TDAB electrophoresis permitted us to obtain a much better resolution of several peptides of the mitochondrial f 0 -f 1 -ATPase, especially for the α and β subunits and for the oligomycin sensitivity conferring protein (OSCP). The differences between the two electrophoretic profiles have been used to devise a new technique of two-dimensional electrophoresis using successively anionic and cationic detergents. This method could be very useful in the case of membrane proteins, which are generally soluble only in the presence of powerful ionic detergents. It has been particularly successful in resolving the small peptides of the f 0 -f 1 -ATPase which were difficult to differentiate by other techniques in one- or two-dimensional polyacrylamide gel electrophoresis.

关键词： Polyacrylamide gel electrophoresis Detergent Cationic detergent Anionic detergent Membrane protein f 0 -f 1 -ATPase (Pig heart mitochondria) SDS sodium dodecyl sulfate TDAB tetradecyltrimethylammonium bromide Bis N,N′- methylene(bis)-acrylamide TEMED N,N,N′,N′- tetramethylethylene diamine ATPase adenosine-5′-triphosphatase (EC 3.6.1.3) f 1 -ATPase pig heart mitochondrial ATPase prepared according to the procedure of Penin et al. [17] OSCP oligomycin sensitivity conferring protein [27] If 1 natural protein inhibitor of mitochondrial f 1 -ATPase [25]

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DIffERENTIAL INHIBITION Of f0f1-ATPASE-CATALYZED REACTIONS IN BOVINE-HEART SUBMITOCHONDRIAL PARTICLES BY ORGANOTIN COMPOUNDS

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第1期766卷 209-214页

作者： EMANUEL, EL CARVER, MA SOLANI, GC GRIffITHS, DE UNIV WARWICK DEPT CHEM & MOLEC SCI COVENTRY CV4 7AL W MIDLANDS ENGLAND INST BIOL CHEM BOLOGNA ITALY

Preincubation of coupled submitochondrial particles with low concentrations of triorganotin compounds results in complete inhibition of the oligomycin-sensitive ATPase activity without any significant effect on the rate of succinate-driven ATP synthesis. The residual ATP synthetic activity is inhibited by oligomycin and uncouplers. The differential inhibition of ATP synthesis and hydrolysis by the triorganotin compounds examined suggests that the 2 processes are not ''mirror images'' of each other, but that they occur through different routes and that the f1f0-ATPase is at least bifunctional.

关键词： f 0 f 1 -ATPase inhibition Organotin (Bovine heart) f 0 and f 1 , components of the mitochondrial ATPase system Hepes, 4-(2-hydroxyethyl)-1-piperazineethane-sulphonic acid

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INCREASE IN CELL-SURfACE N-ACETYLGLUCOSAMINIDE BETA-(1-]4)GALACTOSYLTRANSfERASE ACTIVITY WITH RETINOIC ACID-INDUCED DIffERENTIATION Of f9 EMBRYONAL CARCINOMA-CELLS

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fEBS LETTERS 1984年第2期168卷 222-226页

作者： NAKHASI, HL NAGARAJAN, L ANDERSON, WB NCI TUMOR IMMUNOL & BIOL LABBETHESDAMD 20205

Exposure of f9 cells to all-trans-retinoic acid over a period of 6 days resulted in 4-fold induction of cell surface N-acetylglucosaminide beta (1----4)galactosyltransferase (GT) activity. The retinoic acid-induced GT activity was further enhanced by treatment of the cells with 8-bromo cyclic AMP. The ability of retinoic acid alone, or retinoic acid in combination with 8-bromo cyclic AMP, to induce GT activity was inhibited by both actinomycin D and cycloheximide. These findings indicate that the induction of galactosyltransferase activity noted with differentiation of f9 cells involves de novo synthesis of new enzyme protein.

关键词： f 9 cell Retinoic acid 8-Bromo cyclic AMP Galactosyltransferase activity Differentiation De novo synthesis

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DEMONSTRATION Of 2 BINDING-SITES fOR ADP ON THE ISOLATED BETA-SUBUNIT Of THE RHODOSPIRILLUM-RUBRUM R1f0f1-ATP SYNTHASE

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fEBS LETTERS 1984年第1期178卷 10-14页

作者： KHANANSHVILI, D GROMETELHANAN, Z WEIZMANN INST SCI DEPT BIOCHEMIL-76100 REHOVOTISRAEL

Two ADP binding sites have been demonstrated on the reconstitutively active β-subunit, that was removed from the Rhodospirillum rubrum membrane-bound ATP synthase. One is a high affinity site ( K d = 0.7 μM) that does not require MgCl 2 and is unaffected by it. The second is a low affinity binding site ( K d = 80 μM) that is absolutely dependent on MgCl 2 . for stable binding of ADP to this site, MgCl 2 must be present not only during the binding step but also during the elution-centrifugation step used to separate the β-subunit bound [ 3 H]ADP from the free ligand. When MgCl 2 is removed together with the free ligand [ 3 H]ADP dissociates very rapidly from this second binding site.

关键词： Rhodospirillum rubrum f 0 f 1 -ATPsynthase β-Subunit ADP-binding site MgCl 2 dependence Binding affinity

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COUPLING BETWEEN PROTEOLYTIC PROCESSING AND TRANSLOCATION Of THE PRECURSOR Of THE f1-ATPASE BETA-SUBUNIT DURING ITS IMPORT INTO MITOCHONDRIA Of INTACT-CELLS

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fEBS LETTERS 1984年第1期178卷 161-164页

作者： KOLAROV, J HATALOVA, I Cancer Research Institute Slovak Academy of Sciences Čs.armády 21 812 32 Bratislava Czechoslovakia

The intracellular transport of newly synthesized beta-subunits of the f1-ATPase (beta f1) and of newly synthesized ADP/ATP carrier was followed in isolated rat hepatoma cells. As tested by rapid fractionation of [35S]methionine pulse- and pulse-chase-labeled cells and by sensitivity of labeled polypeptides to externally added protease, the import of beta f1 into mitochondria was strongly inhibited by the additional low concentrations of rhodamine 6G (R6G). In contrast, the import of the ADP/ATP carrier into mitochondria was not affected by the inhibitor. The results imply that the proteolytic processing of the precursor of beta f1 is coupled to its translocation across the mitochondrial membrane.

关键词： Precursor protein Mitochondrial protein transport Processing protease Rhodamine 6G f 1 -ATPase PMSf phenylmethylsulfonyl fluoride βf 1 β-subunit of mitochondrial f 1 -ATPase R6G rhodamine 6G pre-βf 1 precursor of β-subunit of mitochondrial f 1 -ATPase

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论由I_ρ(f;x)向对应Ⅰ(f;x;ρ)的逼近性质的传递

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数学研究与评论 1984年第4期 121-130页

作者：周克成南京大学

本文采用乘子法研究对应奇异积分I_ρ(f;x)与I(f;x;ρ)逼近性质的相互关系,着重讨论I_ρ(f;x)的逼近性质向I(f;x;ρ)的传递问题。文中证明了对应奇异积分能够有同阶的正逼近定理。在一定条件下,分别就0<α<1,0<α<2,α=2... 详细信息

本文采用乘子法研究对应奇异积分I_ρ(f;x)与I(f;x;ρ)逼近性质的相互关系,着重讨论I_ρ(f;x)的逼近性质向I(f;x;ρ)的传递问题。文中证明了对应奇异积分能够有同阶的正逼近定理。在一定条件下,分别就0<α<1,0<α<2,α=2三种情形,证明了对应核的α阶绝对矩之间的精致关系。给出了非周期奇异积分关于Lip函数类的逼近度量的渐近性质。文中建立了对应核型不等式的等价过渡,指出对应奇异积分能够有类同的逆逼近定理。从而若干常用奇异积分的结构与正、逆逼近定理是本文结果的特例。此外,还给出与Ghermanesco非周期奇异积分的正、逆逼近定理和渐近展式。

关键词：定理数学研究函数类附注 f x 奇异积分乘子法逼近性质常数数学

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满足f2k+1+f=0的（1,1）张量场与相应结构

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河南大学学报(自然科学版) 1984年第2期 35-42页

作者：宋鸿藻 ,吴报强

***研究了n维流形M~n上,满足f3+f=0的（1,1）张量场,[1][2]统一了殆复结构与殆接触结构的概念,证明了: 一个n维流形,容许一个（1,1）型,秩为常数k的非0张量场f,使f3+f=0,当且仅当:K为偶数,K=2m,且流形的切丛群为U（m）×O（n-2m）。... 详细信息

***研究了n维流形M~n上,满足f3+f=0的（1,1）张量场,[1][2]统一了殆复结构与殆接触结构的概念,证明了: 一个n维流形,容许一个（1,1）型,秩为常数k的非0张量场f,使f3+f=0,当且仅当:K为偶数,K=2m,且流形的切丛群为U（m）×O（n-2m）。在[3]中,他又和***等研究了满足φ4±φ2=0的（1,1）张量场φ及相应结

关键词：张量场 f=0 f 光滑流形

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MEASUREMENT Of THE RELATIVE ELECTRON-TRANSPORT CAPACITY Of PHOTOSYSTEM-I AND PHOTOSYSTEM-II IN SPINACH-CHLOROPLASTS

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BIOCHIMICA ET BIOPHYSICA ACTA 1984年第2期765卷 186-195页

作者： MCCAULEY, SW TAYLOR, SE DENNENBERG, RJ MELIS, A UNIV CALIF BERKELEY DIV MOLEC PLANT BIOL BERKELEY CA 94720 USA

The ratio of Photosystem (PS) II to PS I electron-transport capacity in spinach chloroplasts was compared from reaction-center and steady-state rate measurements. The reaction-center electron-transport capacity was based upon both the relative concentrations of the PS ***., PS ***. and PS I centers, and the number of chlorophyll molecules associated with each type of center. The reaction-center ratio of total PS II to PS I electron-transport capacity was about 1.8:1. Steady-state electron-transport capacity data were obtained from the rate of light-induced absorbance-change measurements in the presence of ferredoxin-NADP+, potassium ferricyanide and 2,5-dimethylbenzoquinone (DMQ). A new method was developed for determining the partition of reduced DMQ between the thylakoid membrane and the surrounding aqueous phase. The ratio of membrane-bound to aqueous DMQH2 was experimentally determined to be 1.3:1. When used at low concentrations (200 .mu.M), potassium ferricyanide is shown to be strictly a PS I electron acceptor. At concentrations higher than 200 .mu.M, ferricyanide intercepted electrons from the reducing side of PS II as well. The experimental rates of electron flow through PS II and PS I defined a PS II/PS I electron-transport capacity ratio of 1.6:1.

关键词： Electron transport Photosystem I Photosystem II Reaction center (Spinach chloroplast) Chl chlorophyll DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea PS Photosystem P-700 reaction center of PS I Q primary electron acceptor of PS II DMQ 2,5-dimethylbenzoquinone NADP + nicotinamide adenine dinucleotide phosphate DCIP 2,4-dichlorophenolindophenol C f flattening correction factor Tricine N -[2-hydroxy-1,1-bis(hydroxymethyl)ethyl]glycine Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid

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