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PRE-STEADY-STATE PROPERTIES Of BOVINE HEART MITOCHONDRIAL ATPASE - A NUCLEOTIDE-DEPENDENT H+ BURST

BIOCHIMICA ET BIOPHYSICA ACTA 1985年第2期808卷 280-287页

作者： DAGGETT, SG SCHUSTER, SM UNIV NEBRASKA SCH BIOL SCI LINCOLN NE 68588 USA

The transient kinetics of bovine heart mitochondrial ATPase (f1) depleted of loosely bound nucleotides were observed. The activation process which was shown as a lag time before steady-state hydrolysis observed previously was preceded by a proton burst when f1 was stripped of its loose nucleotides. 5''-Adenylylimidodiphosphate (Ado PP[NH]P) or MgATP binding is shown to cause proton release. Maximum proton release per f1 free of loosely bound nucleotides is observed with MgATP. Modification with NBD-CL [7-chloro-4-nitrobenz-2-oxo-1,3-diazole] of f1 that was nucleotide-depleted eliminated the proton burst, which suggests that the modified tyrosine (i.e., in the catalytic subunit) is directly involved in the release of protons.

关键词： f 1 -ATPase Nucleotide binding Chemical modification ATP hydrolysis (Bovine heart, Kinetics) Ado PP [NH] P 5′-adenylylimidodiphosphate NBD-CL 7-chloro-4-nitrobenzo-2-oxo-1,3-diazole Bes 2-[bis(2-hydroxyethyl)amino]ethanesulfonic acid

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A QUANTITATIVE, IMMUNOCHEMICAL DEMONSTRATION Of THE POSTNATAL-DEVELOPMENT Of NEUROTENSIN IN THE MEDIAL PREOPTIC AREA Of THE RAT

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DEVELOPMENTAL BRAIN RESEARCH 1985年第1-2期18卷 241-250页

作者： WOLfSON, B MANNING, RW DAVIS, LG BALDINO, f DUPONT CO DEPT CENT RES & DEVGLENOLDENPA 19036

The postnatal ontogeny of neurotensin (NT) in the medial preoptic area (MPO) of the rat was quantitatively investigated using immunocytochemical and neurochemical techniques. On postpartum day 0, NT-like immunoreactive neurons could not be identified within the MPO by peroxidase-antiperoxidase histochemistry and, in fact, did not appear until 9 days postpartum. After this age, the number of NT cells in the MPO increased ontogenetically, as did concentrations of NT in MPO extracts measured by radioimmunoassay and enzyme-linked immunosorbent assay. In organotypic MPO explants derived from neonatal rats, NT-like immunoreactivity was detected only after the explants were maintained 14 days in vitro, after which, the number of NT-like immunoreactive somata increased as the time in vitro was extended. These findings indicate that a substantial amount of differentiation occurs postnatally in the MPO both in vivo and in vitro and, more specifically, that the ontogenetic expression of NT may be an intrinsic property of the MPO.

关键词： neurotensin ontogeny immunochemistry neonates medial preoptic area (MPO) enzyme-linked immunosorbent assay (ELISA) radioimmunoassay (RIA) ac anterior commissure BAC bed nucleus of the anterior commissure BST bed nucleus of the stria terminalis BST PO preoptic extension of the bed nucleus of the stria terminalis DBB diagonal band of Broca v DBB vertical limb of the diagonal band of Broca f fornix LPO lateral preoptic area LS v ventral lateral septal nucleus LV lateral ventricle MPO medial preoptic nucleus MnPO median preoptic nucleus oc optic chiasm ot optic tract SC suprachiasmatic nucleus SHy septohypothalamic nucleus SO supraoptic nucleus 2n optic nerve III 3rd ventricle

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ACUTE EffECTS Of AEROSOLIZED METAPROTERENOL ON BREATHING PATTERN Of PATIENTS WITH SYMPTOMATIC BRONCHIAL-ASTHMA

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JOURNAL Of ALLERGY AND CLINICAL IMMUNOLOGY 1985年第2期76卷 166-172页

作者： TOBIN, MJ BIRCH, S JENOURI, G SACKNER, MA MT SINAI MED CTR DEPT MED DIV PULM DIS 4300 ALTON RD MIAMI BEACH FL 33140 USA

The effect of 2 sequential puffs of metaproterenol (650 .mu.g each puff), delivered with an auxillary aerosol delivery system, on the breathing pattern of patients with symptomatic bronchial asthma who were monitored noninvasively with respiratory inductive plethysmography was studied. Particular attention was directed to respiratory center drive as reflected by mean inspiratory flow and minute ventilation. Both these components were elevated in the 8 patients whose mean fEV1.0 [forced expiratory volume in 1 s] was 1.43 l (45% predicted normal). Two puffs of metaproterenol produced a maximal increase over baseline in fEV1.0 of 50 .+-. 25% (SD), whereas no change took place in fEV1.0 with placebo administration. This dose of metaproterenol did not alter heart rate or blood pressure throughout the study period of 2 h. Neither mean inspiratory flow, minute ventilation, nor any component of the breathing pattern changed with this partial reversal of bronchoconstriction. These results suggest that the neural mechanism accounting for heightened respiratory center drive in patients with symptomatic bronchial asthma does not wholly depend on bronchoconstriction.

关键词： V T /T 1 Mean inspiratory flow V min Minute ventilation CPAP Continuous positive airway pressure MDI Metered-dose inhaler RIP Respiratory inductive plethysmograph f frequency V T Tidal volume T 1 Inspiratory time T 1 /T TOT fractional inspiratory time RC Rib cage ABD Abdominal MCA Maximum compartmental amplitude fRC functional residual capacity MIf Maximum inspiratory flow PEEP Positive end expiratory pressure

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RESTORATION BY TETRAMETHYL-PARA-PHENYLENEDIAMINE Of PHOTOSYNTHESIS IN DIBROMOTHYMOQUINONE-INHIBITED CELLS Of THE CYANOBACTERIUM SYNECHOCOCCUS SP

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BIOCHIMICA ET BIOPHYSICA ACTA 1985年第1期809卷 74-80页

作者： NANBA, M KATOH, S UNIV TOKYO COLL ARTS & SCI DEPT PURE & APPL SCI MEGURO KU TOKYO 153 JAPAN

Inhibition of electron flow from water to methyl viologen by 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB) was reversed by the addition of N,N,N'',N''-tetramethyl-p-phenylenediamine (TMPD) to cells of a thermophilic cyanobacterium Synechococcus sp. The restored activity is not a simple Photosystem I reaction because it was suppressed by 3-(3'',4''-dichlorophenyl)-1,1-dimethylurea (DCMU). TMPD was also found to restore photosynthetic oxygen evolution in the DBMIB-poisoned cells. That the restored oxygen evolution is coupled with CO2 reduction was indicated by its sensitivity not only to electron-transport inhibitors such as DCMU and phenylmercuric acetate, but also to an inhibitor of CO2 fixation (KCN), uncouplers (carbonyl cyanide michlorophenylhydrazone, methylamine and ammonium chloride) and an energy-transfer inhibitor (N,N''-dicyclohexylcarbodiimide). In the DBMIB-poisoned cells. TMPD accelerated only P-700 reduction, while leaving the reduction kinetics of cytochrome c-553 and cytochrome f inhibited, indicating that P-700 is the site of electron entry from TMPD to Photosystem I. It is concluded that TMPD restores photosynthetic electron transport by accepting electrons in the plastoquinone region and in turn donating them directly to P-700, and thereby bypassing cytochrome c-553 and DBMIB-blocked cytochrome b6-f complexes.

关键词： Photosynthesis Electron transport Cytochrome b 6 - f complex Electron flow inhibitor Electron flow bypass ( Synechococcus sp.) DBMIB 2,5-dibromo-3-methyl-6-isopropyl- p -benzoquinone TMPD N , N , N ′, N ′-tetramethyl- p -phenylenediamine DCMU 3-(3′,4′-dichlorophenyl)-1,1-dimethylurea PS Photosystem PMA phenylmercuric acetate DCCD N , N ′-dicyclohexylcarbodiimide CCCP carbonyl cyanide m -chlorophenylhydrazone Hepes 4-(2-hydroxyethyl)-1-piperazineethanesulphonic acid

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ARACHIDONIC-ACID DERIVATIVES AS MEDIATORS Of ASTHMA

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JOURNAL Of ALLERGY AND CLINICAL IMMUNOLOGY 1985年第2期76卷 259-264页

作者： LEWIS, RA ROBIN, JL HARVARD UNIV SCH MED DEPT MED BOSTON MA 02115 USA BRIGHAM & WOMENS HOSP DEPT RHEUMATOL & IMMUNOL BOSTON MA 02115 USA

Arachidonic acid metabolites generated by the cyclooxygenase pathway and by the various lipoxygenase pathways are produced by both resident pulmonary cells and infiltrating cells from the vascular compartment. The various proinflammatory biologic activities of these naturally occurring compounds include bronchoconstriction, increased vascular permeability, alterations in vasomotor tone, enhanced mucus secretion, and granulocyte adherence and chemotaxis. The leukotrienes derived from the 5-lipoxygenase pathway are particular potent as mediators of inflammation, requiring only nanomolar concentrations for the evocation of their effects. Thus, although a variety of potent cyclooxygenase inhibitors are currently available for anti-inflammatory therapy, therapeutic modalities for the downregulation of leukotriene biosynthesis or efficacy would be highly desirable. Current concepts about the enzymatic cascade in leukotriene generation, the prospects for dietary modification as an adjunct to pharmacotherapeutic intervention, and the implications of specific receptors in leukotrienemediated events are therefore considered.

关键词： HETEs Monohydroxy-arachidonic acid derivatives PGE 2 PGf 2α PGI 2 PGD 2 Prostaglandins E 2 f 2α I 2 D 2 TxA 2 Thromboxane A 2 LTC 4 LTD 4 LTE 4 LTB 4 LTB 5 LTC 5 Leukotrienes C 4 D 4 E 4 B 4 B 5 C 5 , V̇ max30 Maximal mid-expiratory flow rate after inhalation to 60% vital capacity NSAID Nonsteroidal anti-inflammatory drug

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THE STRUCTURE, fUNCTION AND EVOLUTION Of CYTOCHROMES

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PROGRESS IN BIOPHYSICS & MOLECULAR BIOLOGY 1985年第1期45卷 1-56页

作者： MATHEWS, fS Department of Physiology and Biophysics Washington University School of Medicine 660 South Euclid Avenue St. Louis Missouri 63110 U.S.A.

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ATP SYNTHESIS BY ATPASE PROTEOLIPOSOMES fROM THE THERMOPHILIC CYANOBACTERIUM SYNECHOCOCCUS 6716 BY IONOPHORE-INDUCED ELECTRIC POTENTIALS AND PROTON GRADIENTS

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BIOCHIMICA ET BIOPHYSICA ACTA 1985年第2期809卷 236-244页

作者： VANWALRAVEN, HS HAGENDOORN, MJM KRAB, K HAAK, NP KRAAYENHOf, R FREE UNIV AMSTERDAM BIOL LABDE BOELELAAN 10871081 HV AMSTERDAMNETHERLANDS

ATP synthesis driven by low pre-established electric potentials and pH gradients is studied in large ATPase proteoliposomes, prepared from the ATPase complex and native lipids from the thermophilic cyanobacterium Synechococcus 6716. Electric potentials and pH gradients were achieved by valinomycin and nigericin, respectively, in the presence of a K+ gradient across the membrane. External base-pulses were also applied. In this system ATP synthesis driven by valinomycin-induced K+ influx, nigericin-induced internal acidification and by external base-pulses is demonstrated. Electric potentials and pH gradients of equivalent size lead to roughly similar ATP synthesis activities. ATP synthesis is optimal at 80-100 nM valinomycin and at 0.75-1 μM nigericin at the proper pre-set ion gradients. Uncoupler and DCCD inhibit ATP synthesis. Prior activation of the complex by thiol agents or trypsin was not required for synthesis activity. The ATP synthesis rate increases with the size of electric potential or pH gradient. The threshold value of the electrochemical gradient for significant ATP synthesis is about 30 mV. ATP production proceeds for more than 60 min. The generation of ionophore-induced electric potentials and pH gradients have been followed by oxonol VI and intraliposomal Neutral red, respectively. The extent of the absorbance changes of both probes is proportional to the size of electric potential or pH gradient. Ionophore-induced oxonol VI and Neutral red responses are stable for at least 30 min. The results are discussed in terms of membrane permeability and vesicle size.

关键词： Liposomes ATP synthesis, Membrane potential pH gradient ATPase (Cyanobacteria) DCCD N , N ′-dicyclohexylcarbodiimide f 1 , the water-soluble part of the ATPase complex f 0 , the membrane-bound part of the ATPase complex oxonol VI, bis(3-propyl-5-oxoisoxazol-4-yl)pentamethine oxonol S-13,5-chloro-3- t -butyl-2′-chloro-4′-nitrosalicylanilide

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THE NUCLEOTIDE-SEQUENCE AND SOME PROPERTIES Of THE NEUTRAL PROTEASE GENE Of BACILLUS-AMYLOLIQUEfACIENS

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JOURNAL Of BIOTECHNOLOGY 1985年第2期2卷 75-85页

作者： SHIMADA, H HONJO, M MITA, I NAKAYAMA, A AKAOKA, A MANABE, K fURUTANI, Y MITSUI TOATSU CHEM INC CENT RES INST BIOTECHNOL LAB 1900 TOGO MOBARA CHIBA 297 JAPAN

A 1.9 kilobase fragment possessing a gene for extracellular neutral protease of B. amyloliquefaciens was cloned in pNP150. The nucleotide sequence of the fragment revealed putative regions which were considered as a promoter, a ribosome binding site and a transcription terminator as well as a protein coding region. The sequences of -35 and -10 regions of the putative promoter, TTGCAG and TATTAT, resembled the consensus sequences for the promoter recognized by the .sigma.55 RNA polymerase of B. subtilis. The putative ribosome binding site, whose sequence was AAAGGGGG, was completely complementary to the binding sequence of B. subtilis 16S rRNA except for one base. The protein coding region which started at an initiation codon of GTG contained a sequence corresponding to a 221-residue-long stretch of amino acids preceding the N-terminal of the mature enzyme. This suggested that the neutral protease might have a pre-pro structure involved in an unknown function during secretion. The N-terminal amino acid sequence of the mature enzyme secreted by B. subtilis (pNP150) was the same as that of the authentic enzyme produced by B. amyloliquefaciens. The pre-pro structure of B. amyloliquefaciens neutral protease synthesized in B. subtilis was exactly recognized and processed during secretion.

关键词： neutral protease gene Bacillus amyloliquefaciens f DNA amino acid sequence pre-pro structure

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7 COORDINATION IN MOLYBDENUM CHEMISTRY - ANALYSIS Of OLIGONUCLEAR STRUCTURES

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COORDINATION CHEMISTRY REVIEWS 1985年第JUL期65卷 49-85页

作者： MELNIK, M SHARROCK, P SLOVAK UNIV TECHNOL BRATISLAVA DEPT INORGAN CHEM CS-81237 BRATISLAVA CZECHOSLOVAKIA

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MITOCHONDRIAL ADENOSINE-TRIPHOSPHATASE fROM HUMAN-PLACENTA - EffECTS Of ADENYLYL AND GUANYLYL IMIDODIPHOSPHATE

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INTERNATIONAL JOURNAL Of BIOCHEMISTRY 1985年第2期17卷 229-234页

作者： ALEKSANDROWICZ, Z Department of Biochemistry I.B.M. Medical School 80-211 Gdansk Debinki 1 Poland

1. 1. The effects of adenylylimidodiphosphate (AMP-PNP) and guanylylimidodiphosphate (GMP-PNP) on the kinetics of MgATP, MgITP and MgGTP hydrolysis by mitochondrial ATPase (EC 3.6.1.3) from human placenta were studied. 2. 2. AMP-PNP is a noncompetitive inhibitor of hydrolysis of all substrates used, both in the presence and in the absence of the activating staggered − 3 anion. 3. 3. At least two binding sites for AMP-PNP are present in the f 1 . 4. 4. Unlike AMP-PNP, GMP-PNP was shown to be a competitive inhibitor of hydrolysis of all substrates used. 5. 5. The results of the kinetic experiments presented support the alternating three-site mechanism of ATP hydrolysis by mitochondrial ATPase.

关键词： AMP-PNP adenyl-5′-yl imidodiphosphate GMP-PNP guanyl-5′-yl-, -imidodiphosphate ATP adenosine triphosphate GTP guanosine triphosphate ITP insone triphosphate Trisma base N -Tris (hydromethyl)-aminomethane f 1 soluble ATPase or coupling factor 1 f 1 ·AMP-PNP f 1 ·GMP-PNP soluble complex between the enzyme the nucleotide analogue bound at an catalytic site Enzymes ATPase adenosine triphosphate (EC 3.6.1.3) ATP phosphohydrolase lactate dehydrogenase l -lactate NAD + oxidoreductase (EC 1.1.1.27) Pyruvate kinase ATP pyruvate 2- O -phosphotransferase (EC 2.7.1.40)

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