Interaction of the serum amyloid A proteins with phospholipid

The serum amyloid A proteins (SAA) are transported in plasma in association with the high density lipoproteins. We have studied the solution properties of two of the polymorphic forms of SAA, SAA1 and SAA4, and compared the lipid-binding properties of SAA4 to those of the well characterized apolipoproteins, apo-A-I, apo-A-II, and apo-C-III. SAA4 was monomeric at pH 2.9 but considerable self-association was demonstrated at pH 8.2, even in the presence of 1.0 M guanidine HCl. SAA4 differed from the apolipoproteins in its ability to disrupt multilamellar dimyristoylphosphatidylcholine (DMPC) liposomes and generate bilayer discs. Apo-A-I, apo-A-II, and apo-C-III reduced the turbidity of DMPC dispersions at protein:lipid molar ratios of 1:200. SAA4, however, increased turbidity at molar ratios of 1:250 and 1:100 even when preincubated in guanidine HCl before addition to liposomes. Optical density decreased only at ratios of 1:50 and 1:25. At an SAA4:DMPC ratio of 1:50, discoidal particles (long axis, 28.1 nm; short axis, 4.4 nm) were formed which were similar to those produced by apo-C-III. Lipid binding induced changes in SAA4 conformation similar to those observed in the apolipoproteins. The alpha-helical content and intrinsic tryptophanyl fluorescence were increased and quenching of tryptophanyl fluorescence by acrylamide was reduced in the presence of DMPC. In addition, SAA4 as well as the apolipoproteins broadened the range and increased the temperature of the gel-liquid crystal transition temperature of DMPC.     

Tidal flows and sediment budgets for a salt-marsh system,Essex, England

Studies of tidal flows in salt-marsh creeks in Essex, England, show large variations in water velocity during different tidal cycles, particularly between tides below, at, and above marsh level. Water level, velocity and suspended sediment concentration have been monitored at 5-min intervals during 700 tidal cycles during the year March 1982–March 1983, and the data are being used to calculate sediment budgets for the creek system studied. Completed analyses for two of the tidal cycles show a large positive sediment flux. Because of the importance of velocity in controlling total discharge through a creek cross-section, and hence its effect on total sediment movement, we cannot extrapolate from these two below-marsh tides to any general conclusions about marsh erosion or accretion. We use these preliminary data both to demonstrate our methods and to indicate some of the complexities involved in the analysis.Acknowledgements: This work has been supported by the Natural Environment Research Council. We thank the Philip Lake Fund for financial assistance and the Department of Geography, Cambridge University, for much material support. Mr D. J. Fisher kindly gave access to his land, and Mr W. Bailey helped us greatly. We also thank Mr A. St Joseph for his help, Mr M. Diver for practical support, and Dr J. S. Pethick for discussion.     

3. Control of sex expression in conifers

Despite considerable progress in recent years in the promotion of flowering in conifers, relatively little is known about the endogenous control of sex expression or its practical manipulation to aid pollen and crop management in seed orchards. Well defined patterns of sexual zonation exist within the tree crown and shoot, and these appear to be associated with hormonal and possibly nutritional gradients. No firm conclusions, however, are possible regarding the specific role(s) of different PGRs in sex expression, this being as much as a problem of lack of critical study as the complexity of the process itself. Whether a PGR such as gibberellins or auxins, or a cultural treatment that itself may influence endogenous hormone levels, primarily promotes female or male flowering often depends on the position within the crown of the shoot being treated, and its timing in relation to stage of primordia development. Photoperiod also may be involved- short days favoring females and long days males, at least in some species- although its role in controlling sex expression is by no means clear. Practical manipulation of sex expression may be possible through selective crown pruning, different PGR combinations, treatment timing and/or photoperiod control, but the development of such techniques for operational use will require a better basic understanding of the endogenous regulatory mechanisms involved.     

Solid-state 15N NMR of oriented lipid bilayer bound gramicidin A'

Highly oriented samples of lipid and gramicidin A' (8:1 molar ratio) have been prepared with the samples extensively hydrated (approximately 70% water v/w). These preparations have been shown to be completely in a bilayer phase with a transition temperature of 28 degrees C, and evidence is presented indicating that the gramicidin is in the channel conformation. An estimate of the disorder in the alignment of the bilayers parallel with the glass plates used to align the bilayers can be made from the asymmetry of the nuclear magnetic resonances (NMR). Such an analysis indicates a maximal range of disorder of +/- 3 degrees. Uniformly 15N-labeled gramicidin has been biosynthesized by Bacillus brevis grown in a media containing 15N-labeled Escherichia coli cells as the only nitrogen source. When prepared with labeled gramicidin, the oriented samples result in high-resolution 15N NMR spectra showing 12 resonances for the 20 nitrogen sites of the polypeptide. The frequency of the three major multiple resonance peaks has been interpreted to yield the approximate orientation of the N-H bonds in the peptide linkages with respect to the magnetic field. These bond orientations are only partially consistent with the extant structural models of gramicidin.     

Identification of a complex of the three forms of the rat liver asialoglycoprotein receptor

We have generated antibodies against synthetic peptides which represent the carboxyl terminus of either the major, or the two minor, forms of the rat hepatic lectin which recognizes galactose-terminated glycoproteins (asialoglycoproteins). The antibodies were shown to be specific for the form of the lectin containing the immunizing peptide sequence by the following: reaction with purified lectin after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoprecipitation of sodium dodecyl sulfate-denatured lectin, immunoprecipitation of lectin synthesized in vitro. These antibodies, however, precipitated all three rat hepatic lectin forms from nonionic detergent extracts of hepatocytes labeled with 125I via the lactoperoxidase catalyzed technique. A similar result was obtained if antibody was bound to intact cells prior to solubilization with detergent and collection of the immune complexes. We conclude that at least the plasma membrane-associated fraction of the rat hepatic lectin forms exists as a heterotypic complex.