Stereochemistry and size of sugar head groups determine structure and phase behavior of glycolipid membranes: densitometric, calorimetric, and X-ray studies

The role carbohydrate moieties play in determining the structure and energetics of glycolipid model membranes has been investigated by small- and wide-angle X-ray scattering, differential scanning densitometry (DSD), and differential scanning microcalorimetry (DSC). The dependence of a variety of thermodynamic and structural parameters on the stereochemistry of the OH groups in the pyranose ring and on the size of the sugar head group has been studied by using an homologous series of synthetic stereochemically uniform glyceroglycolipids having glucose, galactose, mannose, maltose, or trimaltose head groups and saturated ether-linked alkyl chains with 10, 12, 14, 16, or 18 carbon atoms per chain. The combined structural and thermodynamic data indicate that stereochemical changes of a single OH group in the pyranose ring can cause dramatic alterations in the stability and in the nature of the phase transitions of the membranes. The second equally important determinant of lipid interactions in the membrane is the size of the head group. A comparison of lipids with glucose, maltose, or trimaltose head groups and identical hydrophobic moieties has shown that increasing the size of the neutral carbohydrate head group strongly favors the bilayer-forming tendency of the glycolipids. These experimental results provide a verification of the geometric model advanced by Israelachvili et al. (1980) [Israelachvili, J. N., Marcelja, S., & Horn, R. G. (1980) Q. Rev. Biophys. 13, 121-200] to explain the preferences lipids exhibit for certain structures. Generally galactose head groups confer highest stability on the multilamellar model membranes as judged on the basis of the chain-melting transition. This is an interesting aspect in view of the fact that galactose moieties are frequently observed in membranes of thermophilic organisms. Glucose head groups provide lower stability but increase the number of stable intermediate structures that the corresponding lipids can adopt. Galactolipids do not even assume a stable intermediate L alpha phase for lipids with short chain length but perform only Lc----HII transitions in the first heating. The C2 isomer, mannose, modifies the phase preference in such a manner that only L beta----HII changes can occur. Maltose and trimaltose head groups prevent the adoption of the HII phase and permit only L beta----L alpha phase changes. The DSD studies resulted in a quantitative estimate for the volume change associated with the L alpha----HII transition of 14-Glc. The value of delta v = 0.005 mL/g supports the view that the volume difference between L alpha and HII is minute.(ABSTRACT TRUNCATED AT 400 WORDS)     

Ni(II) and Ni(I) forms of pentaalkylamide derivatives of cofactor F430 of Methanobacterium thermoautotrophicum.

A series of pentaalkylamide forms of F430 and of its 12,13-diepimer have been generated and characterized. Carbodiimide-assisted N-hydroxysulfosuccinimide activation of all five peripheral carboxylates of the F430 macrocycle allows nucleophilic attack by a number of primary amines (RNH2, R- = CH3-, CH3CH2-, CF3CH2-, CH3(CH2)3-) generating the pentaalkylamide derivatives. The identity of each derivative has been verified by fast-atom bombardment mass spectrometry (FAB-MS). The solubility of these derivatives in aprotic organic solvents varies as the amine alkyl substituent (R-) is changed. Electrochemical measurements have shown that the Ni(II/I) reduction potentials in N,N-dimethylformamide (DMF) are approximately -1 V (Ag/AgCl). Reduction by sodium amalgam in THF generates the Ni(I) form of the F430 diepimer pentabutylamide. The visible and EPR spectra of this Ni(I) species are very similar to the corresponding spectra of Ni(I) F430M (Jaun, B. and Pfaltz, A. (1986) J. Chem. Soc. Chem. Commun. 1327-1329.).     

Enhancement of specific nitrogenase activity inAzospirillum brasilense andKlebsiella pneumoniae,inhibition inRhizobium japonicum under air by phenol

Specific nitrogenase activity inAzospirillum brasilense ATCC 29145 in surface cultures under air is enhanced from about 50 nmol C₂H₄·mg protein^-1·h^-1 to 400 nmol C₂H₄ by the addition of 1 mM phenol. 0.5 and 2 mM phenol added increase the rate 5-fold and 4-fold. This enhancement effect is observed only between 2 and 3 days after inoculation, with only a small reduction of the growth of the cells by the phenol added. In surface cultures under 1% O₂, nitrogenase activity is slightly reduced by the addition of 1–0.01 mM phenol. Utilization of succinate is enhanced during the period of maximum enhancement of nitrogenase activity by 60% by addition of 1 mM phenol. The cells did not produce¹⁴CO₂ from [U-¹⁴C] phenol, neither in surface cultures nor in liquid cultures and less than 0.1% of the phenol was incorporated into the cells. A smaller but significant enhancement of nitrogenase activity by about 100% in surface cultures under air was found withKlebsiella pneumoniae K 11 after addition of 1 mM phenol. However, inRhizobium japonicum 61-A-101 all phenol concentrations above 0.01 mM reduced nitrogenase activity. With 1 mM phenol added activity was reduced to less than 10% with no effect on the growth in the same cultivation system. With thisRhizobium japonicum strain significant quantities of phenol (25 mol in 24 h by 2·10¹² cells) were metabolized to¹⁴CO₂, with phenol as sole carbon source. WithAzospirillum brasilense in liquid culture under 1% and 2% O₂ in the gas phase, no enhancement of nitrogenase activity by phenol was noticed.     

The relationship between split-line orientation and in vivo bone strain in galago (G. crassicaudatus) and macaque (Macaca mulatta and M. fascicularis) Mandibles

There is still disagreement concerning the functional significance of split-line patterns in bone. This study was undertaken to reexamine the mechanical forces hypothesis for split-line formation by comparing split-line patterns with in vivo mandibular bone strain patterns. The relationship between split-line orientation and in vivo stress and strain patterns on macaque and galago mandibles was examined during jaw opening and the power stroke of mastication and incision. An attempt was made to relate split-line orientation to the direction of tensile stress and strain along the primate mandible. In addition, we also investigated the alternative possibility that split-line orientation is related to the direction of low stresses (and strains) on the primate mandible. The results of this study showed that there was no consistent relationship between split-line orientation and the principal strains or stresses. Thus, split-lines did not run consistently in the direction of high or low stress and strain. Therefore, we have concluded that split-line orientation provides little useful information for inferring patterns of stress and strain in bone.     

IF-3 crosslinking to Escherichia coli ribosomal 30 S subunits by three different light-dependent procedures: Identification of 30 S proteins crosslinked to IF-3—Utilization of a new two-stage crosslinking reagent, p-nitrobenzylmaleimide

We here report the results of using three light-dependent procedures for crosslinking IF-3 to 30 S proteins within an IF-3·30 S complex. In the first procedure, employing FMN as a photosensitizer, protein S12 is found to be the only major crosslinked protein. In the second procedure, IF-3 is first reacted with the new two-stage crosslinking reagent, p-nitrobenzylmaleimide (PNBM), and the PNBM—IF-3·30 S complex is irradiated. The major crosslinked proteins are S3 > S2, S12, S18. Small amounts of crosslinked S11 and S21 are also found. In the third procedure, the IF-3·30 S complex is reacted with PNBM and then irradiated. The major crosslinked proteins are S12 > S3 > S11 and small amounts of crosslinked S1, S13, and S21 are also found. These results are compared with results obtained by others using different crosslinking procedures and are used to discuss the Lake and Kahan model (J. A. Lake and L. Kahan, 1975, J. Mol. Biol., 99, 631–644, and J. A. Lake, 1978, in Advanced Techniques in Biological Electron Microscopy II, Koehler, J. K., ed., pp. 173–211, Springer-Verlag, Berlin) for IF-3 binding to 30 S subunits.     

FITC-labeled lectins and calcofluor white ST as probes for the investigation of the molecular architecture of cell surfaces. Studies on conjugatophycean species

Summary In a screening program with 7 FITC-labeled lectins as probes, ConA receptors were identified in all of the 28 members of theConjugatophyceae, being under investigation. In nearly all of them RCA₁₂₀ receptors, too, are expressed. In 3 species only, PNA receptors, and in 2 species UEA receptors have been detected. No binding of DBA, SBA, and WGA was observed. The receptors for ConA, RCA₁₂₀, and UEA were shown to be associated with different molecules. Each lectin exhibits a unique and specific binding pattern, both chemically, as well as with regard to the topographic distribution on cell surfaces. While ConA receptors predominantly are associated with constituents of the cell wall, RCA₁₂₀ receptors mostly form part of the surrounding mucilage; the same holds for UEA receptors. Besides a variability of topographic distribution and species-to-species variation, a cell-to-cell variation exists in many species, suggesting that the expression of a lectin receptor is due to the developmental state of the cell and/or depends on external stimuli. In conclusion, we may point out, that FITC-labeled lectins turned out to be extremely useful probes for the investigation of the molecular architecture of cell walls. Calcofluor white ST binding to fibrillar polysaccharides (most probably cellulose) was shown to be inhibited by external incrustations of the cell wall. One species does not show any reaction with calcofluor white ST at all.     

In situ studies on crassulacean acid metabolism in Sedum acre L. and Sedum mite Gil

Summary CO₂ exchange, the diurnal variations in the levels of malic, citric and isocitric acid, and the labelling pattern after ¹⁴CO₂ fixation were measured in Sedum acre and Sedum mite growing in situ. As predicted from laboratory experiments, drought changed the gas exchange pattern from a C₃ type to a crassulacean acid metabolism (CAM) type. This shift correlated with the development of a diurnal rhythm in the malic acid content. The results of ¹⁴CO₂ pulse-chase experiments suggest that in well-watered plants a CAM pattern of carbon flow already exists; hence water stress might enhance latent CAM rather than induce it. The in situ CAM performance by the Sedum species appeared to be highly susceptible to modulation by season and external factors, particularly light and temperature.CAM did not substantially contribute to total carbon gain in S. acre and S. mite. During most of their lifecycles the plants grow under conditions that favour CO₂ uptake by the C₃ pathway rather than by CAM. Hence, despite a capability to feature CAM, the ¹³C values found in S. acre and S. mite are those of C₃ plants.Abbreviations CAM Crassulacean Acid Metabolism - PEP-C Phosphoenolpyruvate-Carboxylase - DW Dry weight Dedicated to Prof. Dr. Dr. h.c. M. Evenari on the occasion of his 75th birthday and to Dr. K.F. Springer     

Your son is cured now; You may take him home

While responding to a question on medical errors in an in-depth interview, a physician told a story about a medical error. The story revealed his silent involvement in the evolution of the error. His response is presented as a text requiring interpretation. Fine details of his manner of speaking are displayed in order to disclose what he said and what it meant. In interpreting the text, phenomenological and sociolinguistic methods are used.