Direct evidence for co-localization of adenylate cyclase,dopamine-β-hydroxylase and cytochrome b562 to bovine chromaffin granule membranes

Adenylate cyclase, dopamine-β-hydroxylase and cytochrome b₅₆₂ have been found to co-equilibrate on equilibrium sucrose gradients of lysed chromaffin granule membranes from bovine adrenal medulla. Peak activities for these enzymes, as well as maximum membrane protein concentration, were found to coincide at d = 1.11 gm cm^?3, and the ratios of adenylate cyclase to both dopamine-β-hydroxylase and cytochrome b₅₆₂ were constant across the entire peak. Adenylate cyclase activity has been reported previously to co-purify with chromaffin granule membranes, and we conclude, on the basis of these new data, that adenylate cyclase is also an intrinsic granule membrane enzyme.     

ATP-stimulated transmitter release and cyclic amp synthesis in isolated chromaffin granules

ATP stimulates chromaffin granules from the bovine adrenal medulla to release epinephrine and specific soluble proteins. ATP analogs substituted in the β-γ-position with either nitrogen or carbon were also found to be effective at inducing release from isolated chromaffin granules. However, an ATP analog substituted at the α-β position with carbon was strongly inhibitory. Cyclic AMP was also found to be synthesized by isolated chromaffin granules under release conditions. ATP analogs were effective as substrates for adenylate cyclase in the same order as their efficiency for inducing release from vesicles. Hydrolysis at the β-γ linkage of ATP therefore is probably not necessary for release; however, hydrolysis at the α-β position may be important in the release process. Cyclic AMP may be produced and play a regulatory role in this event.     

Effect of oxygen on the cyanobacterium Oscillatoria limnetica

Oscillatoria limnetica grown photoautotrophically under aerobic or anaerobic conditions contained a single superoxide dismutase (SOD) of identical electrophoretic mobility in both cases. Its activity was cyanide resistant and H₂O₂ sensitive, implicating Fe-SOD. The enzyme level was high in aerobically and low in anaerobically growing cells. Anaerobically grown cells were more sensitive than aerobic to photooxidation, as expressed by bleaching of phycocyanin and disintegration of the trichomes.Abbreviations DCMU 3-(3,4-dichlorophenyl)-1,1-dimethylurea - SOD superoxide dismutase     

METABOLIC PATTERNS IN THREE TYPES OF PHAGOCYTIZING CELLS

Some chemical and metabolic characteristics of polymorphonuclear leukocytes and monocytes from peritoneal exudates of the guinea pig, and of alveolar macrophages from the same animal, have been compared. Changes in the metabolic patterns of these three types of cell have been followed during the act of phagocytosis. The effect of conventional inhibitors of metabolism, and of anaerobiosis on the phagocytic ability of each of the three cell types mentioned has also been determined. From these studies it was found that alveolar macrophages depend to a considerable degree upon oxidative phosphorylation to provide energy for phagocytosis. The other two types of cell depend only on glycolysis as the source of metabolic energy for that function. In some experiments aimed at obtaining information on the possible role of complex lipids in the function of the cell membrane, it was noted that phagocytosis stimulated the incorporation of inorganic phosphate-P³² into the phosphatides of both types of cell from peritoneal exudates—whether these were free-swimming or adherent to a surface. This phenomenon has not yet been detected in the case of alveolar macrophages.     

Biosynthesis of chlorophyll P-680 of photosystem II in greening leaves of plants adapted to heat shock

In etiolated pea and maize leaves illuminated after incubation at 38 degreesC, a new dark reaction was shown manifested in the bathochromic shift of spectral bands and accompanied by esterification of the product of protochlorophyllide photochemical reduction--Chld 684/676: Chld 684/676 --> Chl 688/680. After completion of the reaction a rapid (20-30 sec) quenching of the fluorescence of the reaction product (Chl 688/680) was observed. The reaction Chld 684/676 --> Chl 688/680 is inhibited under anaerobic conditions and in the presence of cyanide; the reaction accompanied by Chl 688/680 fluorescence quenching is not observed in pea mutants with impaired function of photosystem II reaction centers. The spectral properties of the formed Chl form with the absorption maximum at 680 nm, fluorescence quenching, and simultaneous synthesis of pheophytin suggest that the reaction is connected with the chlorophyll of photosystem II reaction center--P-680.     

COMPARATIVE REPRODUCTIVE BIOLOGY OF ALPINE BIENNIAL AND PERENNIAL GENTIANS (GENTIANA: GENTIANACEAE) IN CALIFORNIA

Reproductive characteristics of three sympatric species of Gentiana exhibiting perennial and non-perennial life histories were studied in alpine meadows of the White Mountains of California during three consecutive years. High fruit and seed set and the production of a relatively large number of seeds characterized the alpine biennials, Gentiana tenella and G. prostrata. In contrast, fruit and seed set were considerably reduced and yearly seed production was relatively low in the alpine perennial, G. newberryi. Successful seed production in the biennial species was a result of low rates of flower predation, self-pollination, and the allocation of a comparatively high proportion of biomass to reproductive structures. Seed production in the perennial species was limited by high rates of flower predation, insufficient amounts of pollen reaching stigmas, an unusually late flowering period, and by the relatively small proportion of biomass allocated to reproduction.     

Probing Structural Elements of Thermal Stability in Bacterial Oligomeric Alcohol Dehydrogenases. I. Construction and Characterization of Chimeras Consisting of Secondary ADHs from Thermoanaerobacter brockii and Clostridium beijerinckii

Summary Two tetrameric secondary alcohol dehydrogenases (ADHs), one from the mesophileClostridium beijerinckii (CBADH) and the other from the extreme thermophileThermoanaerobacter brockii (TBADH), share 75% sequence identity but differ by 26°C in thermal stability. To explore the role of linear segments of these similar enzymes in maintaining the thermal stability of the thermostable TBADH, a series of 12 CBadh and TBadh chimeric genes and the two parental wild-type genes were expressed inEscherichia coli, and the enzymes were isolated, purified and characterized. The thermal stability of each chimeric enzyme was approximately exponentially proportional to the content of the amino acid sequence of the thermophilic enzyme, indicating that the amino acid residues contributing to the thermal stability of TBADH are distributed along the whole protein molecule. It is suggested that major structural elements of thermal stability may reside among the nine discrepant amino acid residues between the N-terminal 50-amino acid residues of TBADH and CBADH.     

Natural-focus infections: the key questions and new vantage points

A short history of the concept of natural focal infections is presented: the idea put forward by D. K. Zabolotny?, E. I. Pavlovski?'s teaching, 3 stages of its development. A number of fundamental questions and the modern content of the concept are considered. The natural foci of infections are a combination of surface, soil and/or water ecosystems, including the population of the causative agent of infection. In contrast to surface ecosystems, in soil and water ecosystems the hosts of the causative agents of sapronotic infections ae soil invertebrates and hydrobios, in which these agents may circulate in biocenotic trophic chains. The circulation of the causative agents in natural foci is a discrete process; the mechanisms and forms of the existence of pathogenic bacteria during seasonal and prolonged periods between epidemics is considered. Special attention is given to latent (nonculturable) forms of bacteria. The complex character of the status of the causative agents of natural focal infections is discussed.     

An 'Old World' scorpion beta-toxin that recognizes both insect and mammalian sodium channels.

Scorpion toxins that affect sodium channel (NaCh) gating in excitable cells are divided into alpha- and beta-classes. Whereas alpha-toxins have been found in scorpions throughout the world, anti-mammalian beta-toxins have been assigned, thus far, to 'New World' scorpions while anti-insect selective beta-toxins (depressant and excitatory) have been described only in the 'Old World'. This distribution suggested that diversification of beta-toxins into distinct pharmacological groups occurred after the separation of the continents, 150 million years ago. We have characterized a unique toxin, Lqhbeta1, from the 'Old World' scorpion, Leiurus quinquestriatus hebraeus, that resembles in sequence and activity both 'New World'beta-toxins as well as 'Old World' depressant toxins. Lqhbeta1 competes, with apparent high affinity, with anti-insect and anti-mammalian beta-toxins for binding to cockroach and rat brain synaptosomes, respectively. Surprisingly, Lqhbeta1 also competes with an anti-mammalian alpha-toxin on binding to rat brain NaChs. Analysis of Lqhbeta1 effects on rat brain and Drosophila Para NaChs expressed in Xenopus oocytes revealed a shift in the voltage-dependence of activation to more negative membrane potentials and a reduction in sodium peak currents in a manner typifying beta-toxin activity. Moreover, Lqhbeta1 resembles beta-toxins by having a weak effect on cardiac NaChs and a marked effect on rat brain and skeletal muscle NaChs. These multifaceted features suggest that Lqhbeta1 may represent an ancestral beta-toxin group in 'Old World' scorpions that gave rise, after the separation of the continents, to depressant toxins in 'Old World' scorpions and to various beta-toxin subgroups in 'New World' scorpions.