The interrelationship between photosynthetic electron transport, glycolate excretion and amino acid metabolism in the blue-green alga Anacystis nidulans1

Photosynthetic rate was measured at (1) a wavelength of 619nm, which predominantly excites PS II although PS I is alsosignificantly excited; (2) 446 nm, which excites mainly PS I;(3) 687 nm for the excitation of PS I; and (4) the wavelengthcombinations of 619+446 and 619+687 nm for enhancement studies.The observed photosynthetic enhancement was 11 to 17%, whileglycolate excretion into the medium was reduced by approximately33%. The production of the amino acid serine also decreased significantly,15 to 19%. A small but insignificant reduction in the productionof glycine was also observed. We suggest that some glycolatewas consumed in an oxidation process associated with PS I duringphotosynthetic enhancement. In this situation, we assume thatthe supply of electrons from the water-splitting process ofPS II is too low for efficient reduction of NADP. This may partlybe compensated by the oxidation of glycolate in the electrontransport chain of photosynthesis in a process associated withPS I. Since the production of amino acids associated with theglycolate pathway also decreased, we conclude that the productfrom the photooxidation of glycolate connected to PS I was directedout of the glycolate pathway. (Received November 28, 1978; )     

Binding proteins for adenosine 3′:5′-cyclic monophosphate in bovine adrenal cortex

Five peaks of cyclic AMP-binding activity could be resolved by DEAE-cellulose chromatography of bovine adrenal-cortex cytosol. Two of the binding peaks co-chromatographed with the catalytic activities of cyclic AMP-dependent protein kinases (ATP-protein phosphotransferase, EC 2.7.1.37) of type I or type II respectively. A third binding protein was eluted between the two kinases, and appeared to be the free regulatory moiety of protein kinase I. Two of the binding proteins for cyclic AMP, sedimenting at 9S in sucrose gradients, could also bind adenosine. They bound cyclic AMP with an apparent equilibrium dissociation constant (K(d)) of about 0.1mum, and showed an increased binding capacity for cyclic AMP after preincubation in the presence of K(+), Mg(2+) and ATP. The two binding proteins differed in their apparent affinities for adenosine. The isolated regulatory moiety of protein kinase I had a very high affinity for cyclic AMP (K(d)<0.1nm). At low ionic strength or in the presence of MgATP, the high-affinity binding of cyclic AMP to the regulatory subunit of protein kinase I was decreased by the catalytic subunit. At high ionic strength and in the absence of MgATP the high-affinity binding to the regulatory subunit was not affected by the presence of catalytic subunit. Under all experimental conditions tested, dissociation of protein kinase I was accompanied by an increased affinity for cyclic AMP. To gain some insight into the mechanism by which cyclic AMP activates protein kinase, the interaction between basic proteins, salt and the cyclic nucleotide in activating the kinase was studied.     

Isolation and characterization of the dut gene of Escherichia coli. II. Restriction enzyme mapping and analysis of polypeptide products

Restriction endonuclease mapping of previously constructed dut plasmids has been carried out using the enzymes PvuI, PvuII and SacI. Various dut plasmids were also tested in the "maxicell" protein-synthesizing system. They all show two protein bands in common, one of Mr 16000 in agreement with the size previously reported for the purified dUTPase subunit (Shlomai and Kornberg, 1978). With the information obtained the structural gene for dUTPase can be assigned to a 950-bp SacI-PvuII fragment of the E. coli genome. Studies, described in the preceding paper, on the overproduction of dUTPase by bacterial strains carrying different dut plasmids strongly suggest that the dut gene is transcribed in the direction from the SacI site towards the PvuII site and that the SacI site is located within the dut control region. The second protein band observed in the "maxicell" experiments has an Mr of 23500. Its identity is unknown but it may represent a precursor of dUTPase or the product of a separate gene located between dut and pyrE.     

High surface hydrophobicity ofStaphylococcus aureus as revealed by hydrophobic interaction chromatography

Hydrophobic interaction chromatography (HIC) on Octyl Sepharose^R in a column procedure was used to compare the relative surface hydrophobicity ofStaphylococcus aureus reference strains, protein A-negative mutants, and strains isolated from bovine mastitis. High protein A-producing strains (Cowan 1 and clinical isolate SA 17970) showed a higher relative surface hydrophobicity than did strains producing a low amount of protein A. One encapsulatedS. aureus strain (Smith diffuse) did not bind to the gel, while an unencapsulated variant showed binding properties similar to weak protein A-producing strains. Studies onS. aureus strains isolated from bovine mastitis revealed a good correlation between adsorption to Octyl Sepharose and the production of protein A. Results indicate that protein A and probably other surface proteins such as fibronectin-binding protein contribute to the high relative surface hydrophobicity ofS. aureus.     

Thermal Rearrangement of 4-Alkyl-1,2,4-triazoles. Rearrangements in the Crystalline State

Studies of the thermolyses of 4-alkyl substituted 1,2,4-triazoles was reviewed. They were observed to rearrange at 200–350 °C to the corresponding 1-alkylated triazoles. When substituted in the 4-position with aryl- or vinylic substituents the triazoles were inert to thermolysis, contrary to what was observed for the 4-alkyl- and 4-allyl substituted systems. The mechanisms for the reactions were elucidated, e.g., by studies of substituents effects and by kinetic measurements in solution as well as for the neat samples. Reactions in solutions were slow. The rearrangements in melts of the neat triazoles readily proceeded to the products, and were proposed to take place via a series of nucleophilic displacement steps. X-ray crystallographic measurements of selected structures, showed that the interatomic distances and angles between the relevant atoms in these structures, to a large degree resembled the geometry expected for the S_N2-type transition states proposed for the rearrangement mechanism. Thus, thermolyses of a series of triazole structures at temperatures below their melting points, confirmed that rearrangements actually did take place. The “kinetics” of the reactions in the crystalline state were investigated and rate constants and thermodynamic data were correlated with the structural characteristics of the crystals.     

The diet of two species of Isoperla (Plecoptera: Perlodidae) in relation to season,site, and sympatry

The seasonal change in gut contents of nymphs of Isoperla grammatica and I. difformis from six streams in southern Sweden was analysed. Both species had ingested a variety of benthic prey and vegetable matter, predominantly diatoms. Some seasonality was evident with high percentages of diatoms in spring in I. grammatica, and in autumn in I. difformis. The scope of food was larger in the latter species which contained about equal proportions of vegetable matter, chironomids, mayfly, stonefly, and black fly larvae. In I. grammatica plant matter and chironomids dominated strongly, comprising > 50% of the gut contents on an annual basis, > 90 % in spring. While small nymphs of I. difformis contained a low proportion of animal matter, only gradually increasing with size, the nymphs of I. grammatica were carnivorous from very early instars. Both species switched to a temporarily strong utilization of algae in spring. This differed among sites, and appeared to reflect differences in insolation and thus the availability of algae. There was a significant negative relationship between the mean densities of Isoperla nymphs and the proportion of animal material found in the guts of I. grammatica (R ² = 0.86). Considering the density of I. grammatica alone, the relationship was weaker (R ² = 0.56). A positive correlation between predator and prey size was observed. With chironomid prey the size range increased with predator size. With simuliid prey, however, prey size increased with predator size in such a way that it suggests selection rather than just an expanding prey size range. Correlations were stronger and regression coefficients significantly higher for I. grammatica than for I. difformis. We suggest that I. grammatica, which ingests a much wider size range of prey might choose prey of optimal sizes more readily than the more synchronously developing I. difformis. Although the life cycles of the two species are staggered, overlap in size distribution indicates that competition for food could be important in spring. However, observed differences in diet should facilitate coexistence. Gut content differences might in turn be accomplished through microhabitat segregation.     

Comparison of an electrochemiluminescent homogenous immunoassay and an ELISA for monitoring productivity in mammalian cell bioreactors

Summary An Enzyme Linked Immuno Sorbent Assays (ELISA) and an Electro-chemiluminescent Immuno Assay (CIA) are compared for the purpose of monitoring product formation in mammalian cell bioreactors. The ELISA had a relative standard deviation of 10%, compared to 8% for the CIA . The CIA was found to be a fast and accurate alternative to the ELISA. The use of more than one immuno assay format was also shown to provide additional insight into process performance.