A small protein in model membranes: a time-resolved fluorescence and ESR study on the interaction of M13 coat protein with lipid bilayers

Model membranes with unsaturated lipid chains containing various amounts of M13 coat protein in the -helical form were studied using time-resolved fluorescence and ESR spectroscopy. The lipid-to-protein (L/P) ratios used were > 12 to avoid protein-protein contacts and irreversible aggregation leading to -polymeric coat protein. In the ESR spectra of the 12-SASL probe in dioleoyl phosphatidylcholine (DOPC) bilayers no second protein induced component is observed upon incorporation of M13 coat protein. However, strong effects are detected on the ESR lineshapes upon changing the protein concentration. The ESR lineshapes are simulated by assuming a fixed ratio between the parallel (D) and perpendicular (D) diffusion coefficients of 4, and an order parameter equal to zero. It is found that increasing the protein concentration from L/P to L/P 15 results in a decrease of the rotational diffusion coefficient D from 3.4 × 10⁷ to 1.9 × 10⁷ s^–1. In the time-resolved fluorescence experiments with DPH-propionic acid as a probe, it is observed that increasing the M13 coat protein concentration causes an increase of the two fluorescent lifetimes, indicating an increase in bilayer order. Analysis of the time-resolved fluorescence anisotropy decay allows one to quantitatively determine the order parameters P₂ and P₄, and the rotational diffusion coefficient D of the fluorescent probe. The order parameters P₂ and P₄ increase from 0.34 to 0.55 and from 0.59 to 0.77, respectively, upon adding M13 coat protein to DOPC bilayers with an L/P ratio of 35. The rotational diffusion coefficient D of the DPH-propionic acid probe decreases on incorporating M13 coat protein, in accordance with the ESR results. It is concluded that M13 coat protein in the -monomeric state is not able to produce a long living lipid boundary shell and consequently an immobilization of the lipids. An overall effect on the lipids is induced, resulting in a reduction in the dynamics and an increase in average lipid order. The hydrophobic region of M13 coat protein is proposed to perfectly match the lipid bilayer, resulting in a relatively small distortion of the bilayer structure of the lipid system.     

Phytoplankton primary production and nutrients in the Oosterschelde (The Netherlands) during the pre-barrier period 1980–1984

Phytoplankton primary production, nutrient concentrations and turbidity were monitored at three stations in the Oosterschelde during 1980–1984 as part of an ecosystem study.From comparisons of dissolved nutrient ratios with the nutrient requirements of phytoplankton, and of ambient nutrient concentrations with half-saturation constants for nutrient uptake by natural phytoplankton populations it was concluded that silicate was a limiting nutrient for diatoms after the spring bloom until the end of the summer. Dissolved inorganic nitrogen and phosphate were not considered to be limiting to phytoplankton growth.In general, the phytoplankton growing season started during the first fortnight of April and ended at the end of September. Column production in the whole Oosterschelde varied between 201 and 540 g C m^–2 yr^–1 and was, on average, 25% higher in the western part than in the eastern part. Basin production in the Oosterschelde varied between 120 and 466 g C m^–2 yr^–1 and was, on average, 55% higher in the western part than in the eastern part; this difference could be explained by differences in the ratio of euphotic depth to mean depth of the compartments.Estimated carbon-specific growth rates in the eastern part varied between < 0.1 and 3 d^–1 and in the western part between < 0.1 and 1 d^–1. This difference could be explained by the great differences in depth of the compartments. Carbon-specific growth rates are discussed in relation to phytoplankton loss rates. It is suggested that in the eastern part sedimentation must be an important sink for phytoplankton.Communication no. 473 of the Delta Institute for Hydrobiological Research, Yerseke, The Netherlands.     

Correlation of rotifer associations in a chain of lakes fed by reclaimed sewage

The quantitative composition of the rotifer fauna of seven interconnected lakes located in southern California was studied in a preliminary investigation. The lakes are fed by reclaimed sewage, and the water quality increases rapidly from the first lake, barely able to support life, to a series of lovely lakes stocked with fish and utilized for recreation. The Shannon index of rotifer population diversity showed an appropriate increase. Up to 24 – 28 different rotifer taxa in each lake provided a rich community dominated by Brachionids, Keratella and Trichocerca. The diversity and quantity of the rotifer association then declined again in the last lake. Comparison of the lakes using the Bray and Curtis index of similarity and cluster analysis supported the picture of a lake-chain evolution, a biotic development showing a maximum curve.     

Studies on the binary and ternary complexes formed by a neurospora glutamate dehydrogenase and its substrates

The NADP⁺ specific glutamate dehydrogenase from wild-type $\text{Neurospora crassa}$ forms a stable binary complex with NADPH. This can combine with L-glutamate, α-ketoglutarate or the substrate analogue D-glutamate to form ternary complexes which can be distinguished by their different fluorescence properties. The affinity of the enzyme for NADPH diminishes with increases in pH or ionic strength of the solution. Experimental data obtained using modified glutamate dehydrogenases from mutant strains of $\text{N. crassa}$ suggest that the reduced-coenzyme binding sites observed fluorimetrically are the same as those observed by enzyme kinetics.     

Structure--activity relationships of chemical inducers of carotenoid biosynthesis

Fifteen amines having a profound effect on carotenogenesis in Marsh seedless grapefruit are reported. The compounds fall into three series: Et₂N(CH₂)_nMe (n = 4–8), Et₂N(CH₂)_nPh (n = 1–5), and Et₂NCH₂CH₂OC₆H₄R (R=H, p-Me, p-Et, p-iso-Pr, p-tert-Bu), There was up to an 11-fold increase in the total carotene content. Lycopene, not normally accumulated, became a major pigment. The inducing ability of the amines on carotenoid biosynthesis is correlated with the octanol-water partition coefficient. The mode of action appears to be similar to that of 2-(4-chlorophenylthio)triethylamine hydrochloride.     

Assessing Methods for Developing Phosphorus Desorption Isotherms from Soils using Anion Exchange Membranes

Developing desorption isotherms for inorganic phosphorus (P) is a time-consuming and non-standardized procedure. Anion exchange membranes (AEMs) have been successfully used in studies of P desorption kinetics and total membrane-desorbable P, but rarely have they been used for developing P desorption isotherms. Our study had two objectives: (1) to evaluate the suitability of using multiple strips of AEMs (termed the Multiple AEM Method) to develop P desorption isotherms; and (2) to compare the Multiple AEM Method with a sequential-extraction approach using AEMs (termed the Sequential AEM Method) to determine if the manner in which AEMs were used would influence the slope of the desorption isotherm, i.e. the partition coefficient. Both methods yielded well-defined, but numerically different desorption isotherms for all levels of sorbed P. However, estimated K _d values among methods were equivalent in the low and medium levels of P sorbed. The Multiple AEM method was quicker than the Sequential AEM method, but both gave similar K _d values in an agriculturally significant range of soil solution concentrations. These methods should be tested on a range of soil type to determine their suitability in developing P desorption isotherms and to move toward method standardization for desorption isotherms.