Phospholipid binding and self-association of the major apoprotein of human and baboon high-density lipoproteins.

The purpose of this study was to establish a relationship between self-association and phospholipid binding of the human and the baboon apoA-I protein. The enthalpy changes on binding dimyristoyl lecithin and lysolecithin to either the human or the baboon native apoA-I protein were measured in a microcalorimeter. An endothermal process, most pronounced for the human apoprotein, was observed at low phospholipid levels. At higher phospholipid to protein ratios the binding was exothermal. Gel filtration experiments on Sephadex G-200 showed that the native apoprotein of both species consists of dimers and tetramers. The baboon native apoA-I protein contained a higher amount of dimers. After preincubation of the apoA-I protein with lysolecithin, the enthalpy changes measured on subsequent binding of dimyristoyl lecithin were shifted towards more exothermal values compared to the curve for the native apoprotein. The amplitude of this shift corresponds to that of the endothermal process observed on binding dimyristoyl lecithin to the native apoprotein. This process was attributed to a phospholipid-induced disaggregation of the apoA-I protein. Gel filtration data showed a decreased extent of aggregation in the apoA-I protein preincubated with lysolecithin. This sample consisted exclusively of dimers. Ultracentrifugal flotation of the complexes formed between the apoA-I protein, and respectively dimyristoyl lecithin and sphingomyelin indicated that preincubation with lysolecithin increased the extent of complex formation. These results suggest that the dimeric form of the apoA-I protein possesses the highest affinity for phospholipids. Any dissociation of higher polymers enhances the phospholipid-binding capacity of the human and the baboon apoA-I protein.     

Ionization behaviour of native apolipoproteins and of their complexes with lecithin. 1. Calorimetric and potentiometric titration of the native apoA-I protein and of the apoA-I protein-dimyristoyl lecithin complex.

The ionization behaviour of native apoA-I protein is compare to that of its complex with synthetic dimyristoyl lecithin in studies using calorimetric, potentiometric and spectrophotometric titration. In the presence of phospholipids, 10 out of 21 lysines together with 22 acidic residues are masked in the complex. All tyrosines remain accessible to titration below pH 13. The apparent ionization enthalpy of the 11 lysine residues is not affected by the presence of phospholipids. These data are consistent with discrete binding sites located in the apoprotein helical segments as suggested by the model of Segrest et al. [FEBS Lett. 38, 247-253 (1974)]. A tentative localisation of lysine, arginine, aspartic acid and glutamic acid residues directly involved in phospholipid binding is suggested, assuming that such helical regions are involved in apoprotein-phospholipid association.     

Thermodynamics of lipid protein associations. Thermodynamics of helix formation in the association of high density apolipoprotein A-I (apoA-I) to dimyristoyl phosphatidylcholine.

The structure and phospholipid-binding properties of human plasma high density apolipoprotein A-I (apoA-I) has been studied at pH 7.4 and 3.1 by microcalorimetry, circular dichroism and density gradient ultracentrifugation. At pH values of 7.4 and 3.1, apoA-I binds to dimyristoyl phosphatidylcholine (DMPC) to form complexes of similar composition (molar ratio of DMPC/apoA-I of 100) and helical content (67%). At pH 7.4, the lipid-protein association is accompanied by an increase in helical content from 58 to 67% and an exothermic enthalpy of binding (deltaHB) of -90 kcal/mol apoA-I. At pH 3.1, the helical content of apoA-I is increased from 48 to 67% on binding to DMPC and the enthalpy of binding was -170 kcal/mol. We suggest that the difference in the enthalpies of binding (-80 kcal/mol) at pH 3.1 compared to 7.4 is due to the greater coil leads to helix transition at the lower pH.     

Effect of prostaglandins F2α and E2 on milk ejection, blood pressure and blood flow through the mammary artery in the cow

The influence of prostaglandins (PG) F₂α and E₂ on milk ejection, mammary artery blood flow and arterial blood pressure was studied in lactating cows. Injections of both PG in the jugular vein or the carotid artery induced milk ejection after a relatively long latency period. The minimal effective dose amounted to 1 to 5 μg and to 100 to 300 μg for PGF₂α and PGE₂ respectively. In several cases with PGF₂α and once with PGE₂ milk ejection was accompanied with a simultaneous increase in blood flow through the mammary artery whereas arterial blood pressure remained unchanged. Both routes of administration showed the same response. It was suggested that the effect of the PG on the bovine myoepithelium is indirect, possibly secondary to a release of oxytocin from the neurohypophysis.     

Ergatoid queens and intercastes in ants: Two distinct adult forms which look morphologically intermediate between workers and winged queens

Summary The term ergatogyne is used in ants to describe permanently-wingless female adults which are morphologically intermediate between workers and winged queens. This definition is ambiguous because there are two distinct categories of ergatogynes: ergatoid queens and intercastes. Both have an external appearance (ocelli and alitrunk structure) which combines traditional queen and worker characters, and thus can be confused if they both function as reproductives — however intercastes in most species cannot reproduce.Ergatoid queens have replaced winged queens in a substantial number of species. They are sometimes externally similar to conspecific workers, especially in various ponerine species which exhibit limited size dimorphism between castes. Ergatoid queens retain the specialized attributes of a reproductive caste, including larger ovaries, and they are always the functional egg-layers in a colony. In contrast, conspecific intercastes represent various graded stages in a series connecting workers and winged queens, and they occur together with the queens. These hybrid phenotypes result from deviations from the normal pattern of caste differentiation during larval development. Intercastes generally lack a spermatheca and have no reproductive function; however they can mate in a few leptothoracine ants, and then reproduce instead of winged queens in a proportion of colonies.     

Trace metals in the benthic habitat of the Maarsseveen Lakes system,The Netherlands

The cadmium, zinc, lead and copper concentrations in benthic invertebrates and sediment were determined during two consecutive winters in the Maarsseveen Lakes system. A sequential extraction procedure was applied to estimate the bioavailability of the trace metals in the sediment. Based on the trace metal analyses of organisms and sediment, it is concluded that the Maarsseveen Lakes system has background levels of cadmium, zinc, lead and copper. As the majority of metals was present in geochemically more stable sediment phases, the sequential extractions provided limited additional information on trace metal bioavailability.     

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.