Evidence for a 13,14-Cis Cycle in Bacteriorhodopsin

We discuss to what extent the vibrational spectra of bacteriorhodopsin that have been observed and assigned by Smith et al. (1, 2) by means of resonance Raman and by Gerwert and Siebert (EMBO (Eur. Mol. Biol. Organ.) J. In press) by means of infrared absorption experiments are in agreement with a photo-cycle of bacteriorhodopsin that involves the sequence BR, IO(all-trans) → K(13,14-cis) → L(13,14-cis) → M(13-cis) → N(13-cis) → O(all-trans). Our discussion is based on a quantumchemical modified neglect of diatomic overlap [MNDO] calculation of the vibrational spectra of the relevant isomers of the protonated retinal Schiff base. In particular, we investigated in these calculations the effects of different charge environments on the frequencies of the relevant C-C single bond stretching vibrations of these isomers.     

Electron Transfer in the Photosynthetic Membrane: Influence of PH and Surface Potential on the P-680 Reduction Kinetics

The primary electron donor P-680 of the Photosystem-II reaction center was photoxidized by a short flash given after dark adaptation of photosynthetic membranes in which oxygen evolution was inhibited. The P-680⁺ reduction rate was measured under different conditions of pH and salt concentration by following the recovery of the absorption change at 820 nm. As previously reported for Tris-washed chloroplasts (Conjeaud, H., and P. Mathis, 1980, Biochim. Biophys. Acta, 590:353-359) a fast phase of P-680⁺ reduction slows down as the bulk pH decreases. When salt concentration increases, this fast phase becomes faster for pH above 4.5-5 and slower below. A quantitative interpretation is proposed in which the P-680⁺ reduction kinetics by the secondary electron donor Z are controlled by the local pH. This pH, at the membrane level, can be calculated using the Gouy-Chapman theory. A good fit of the results requires to assume that the surface charge density of the inside of the membrane, near the Photosystem-II reaction center, is positive at low pH values and becomes negative as the pH increases, with a local isoelectric point ~4.8. These results lead us to propose a functional scheme in which a pH-dependent proton release is coupled to the electron transfer between secondary and primary donors of Photosystem-II. The H⁺/e ratio varies from 1 at low pH to 0 at high pH, with a real pK ~6.5 for the protonatable species.     

Regulation of B-lymphocyte activation, proliferation, and immunoglobulin secretion

Lymphocyte growth and differentiation are controlled by signals resulting from the interaction of antigen and cellular products, such as lymphokines, with specific cell membrane receptors. Resting B lymphocytes can be activated by low concentrations (1-5 micrograms/ml) of antibodies to membrane IgM, which is the B-lymphocyte receptor for antigen. The binding of anti-IgM to B cells causes a rapid increase in intracellular free calcium concentration ([Ca2+]i), in inositol phosphate concentration, and in protein kinase activity. Moreover, the effects of anti-IgM on B cells are mimicked by the combined use of calcium ionophores and phorbol esters. Since phorbol esters activate protein kinase c, this suggests that the increase in [Ca2+]i and in phosphatidylinositol metabolism stimulated by anti-IgM are critical events in B-cell activation. The entry into S phase of B cells stimulated with anti-IgM depends on the action of a T-cell-derived factor designated B-cell stimulatory factor (BSF)-1. This is a 20,000-Da protein which is a powerful inducer of class II major histocompatibility complex molecules. Although an important cofactor for B-cell proliferative responses to anti-IgM, its major locus of action is on resting B cells. B cells stimulated with anti-IgM and BSF-1 do not synthesize secretory IgM. However, if two additional T-cell-derived factors, B151-TRF and interleukin-2, are added to cultures, a substantial proportion of stimulated B cells produce secretory IgM. BSF-1 has also been shown to participate in the "switch" in Ig class expression. Resting B cells cultured with lipopolysaccharide will switch to IgG1 secretion in the presence of purified BSF-1.     

The synaptonemal complex of Meloidogyne nataliei and its relationship to that of other Meloidogyne species

The four synaptonemal complexes (SC) in Meloidogyne nataliei (n=4) have normal, tripartite morphology, although the total width of the SC is only 50 nm. Each SC is attached at both ends to the nuclear envelope and there is no bouquet formation at pachytene. Pairing of homologues is regular but not complete, as there is one region on each bivalent where the SC is not formed. This region may be the fusion point of telomeres of nonhomologous chromosomes, since it is assumed that M. nataliei has been derived from an ancestral strain (n=8) via chromosomal fusions. In each pachytene nucleus there is a nonmembranous, vacuole-like structure of unknown function located in the nucleoplasm adjacent to the nucleolus, and of approximately the same volume as the nucleolus. The fact the SC structure of M. nataliei is strikingly different from that of most Meloidogyne species suggest that M. nataliei may not belong to the same phyletic group as the genus Meloidogyne.     

Comparison of the roles of Ostracods and Cladocerans in regulating community structure and metabolism in freshwater microcosms

Laboratory microcosms were used to compare the effects of the littoral ostracod Cypridopsis vidua and the planktonic cladoceran Daphnia magna on community structure and metabolism. Filter-feeding by cladocerans, both in the presence and absence of ostracods, greatly reduced the abundance of planktonic algae when D. magna reached peak density around day 50; rotifers and euglenids were then limited to flocculent matter on the container bottom. Both net production and community respiration rates decreased as community composition changed. Microcosms containing ostracods as the only microcrustacean showed little reduction in total algal numbers but the otherwise dominant alga, Scenedesmus spp., was replaced by Ankistrodesmus spp. when peak ostracod density was reached around day 100. Rotifers were completely eliminated but euglenids were able to coexist with ostracods. Ostracods impacted community metabolism less than cladocerans, but depressed respiration slightly more than net production.     

In vitro translation of B-cell stimulatory factor-1 in Xenopus laevis oocytes

B-cell stimulatory factor-1 (BSF-1) can be translated in vitro in Xenopus laevis oocytes. This activity is blocked by an antibody to BSF-1. The RNA species coding for BSF-1 activity sediments of approximately 8-9S and is separable from RNA coding for interleukin-2 activity which sediments at approximately 11.5S. Finally, the fact that BSF-1 can be translated in vitro confirms that functions attributed to BSF-1 do not depend on contamination with other biologically active molecules such as phorbol myristate acetate.     

BSF-1 action on resting B cells does not require elevation of inositol phospholipid metabolism or increased [Ca2+]i

B cell stimulatory factor-1 (BSF-1) acts on resting B cells to increase expression of class II major histocompatibility complex (MHC) molecules and to prepare for more prompt entry into S phase in response to anti-IgM and lipopolysaccharide. It also acts as a costimulant, with low concentrations of anti-IgM, to cause resting B cells to synthesize DNA. Unlike anti-IgM, BSF-1 does not cause elevation in inositol phospholipid metabolism or in concentration of intracellular free calcium, nor does it enhance such biochemical responses to anti-IgM. Furthermore, increased expression of class II MHC molecules to BSF-1 is observed when essentially all extracellular calcium is chelated by EGTA, whereas lower concentrations of EGTA completely inhibit increases in class II molecules in response to anti-IgM. These results indicate that BSF-1 effects on resting B cells are not mediated by the inositol phospholipid metabolic pathway.     

Production of BSF-1 during an in vivo, T-dependent immune response

BSF-1, a cytokine produced by some T lymphocyte tumors, has been shown to act with anti-Ig antibodies to stimulate B lymphocyte proliferation, to independently induce resting B lymphocytes to increase their expression of surface Ia antigen, and to induce some activated B lymphocytes to differentiate into IgG1- or IgE-secreting cells. To determine whether BSF-1 might be secreted by normal lymphoid cells in the course of a physiologic immune response, BALB/c mice were injected with an affinity-purified goat antibody to mouse IgD (GaM delta), which induces the generation of a large, polyclonal T-dependent IgG1 response; 4-hr culture supernatants of spleen cells from these mice were prepared, and these supernatants were assayed for BSF-1 activity by analyzing their ability to induce BALB/c nu/nu spleen cells to increase their expression of cell surface Ia in vitro. Culture supernatants of unfractionated spleen cells removed from mice 4 to 8 days after GaM delta antibody injection induced substantial increases in B lymphocyte surface Ia expression; these increases were blocked by a monoclonal anti-BSF-1 antibody. Culture supernatants of spleen cells from untreated BALB/c mice or from untreated or GaM delta antibody-treated BALB/c nu/nu mice induced small to moderate increases in B cell surface Ia expression, and GaM delta antibody itself induced large increases in B cell surface Ia expression; however, these increases were not significantly blocked by a monoclonal anti-BSF-1 antibody. A culture supernatant of T cell-enriched spleen cells from untreated mice induced small increases in B cell surface Ia expression that were inhibited by anti-BSF-1 antibody, as was the larger increase in B cell Ia expression induced by a culture supernatant of T cell-enriched spleen cells from mice sacrificed 3 days after GaM delta injection. On the other hand, T cell-depleted spleen cells from BALB/c mice injected with GaM delta antibody 7 days before sacrifice failed to generate culture supernatants with BSF-1 activity. Supernatants prepared from spleen cells taken from untreated mice or mice treated with GaM delta antibody 1 to 3 days before sacrifice did not block the ability of purified BSF-1 to induce an increase in B cell surface Ia expression, and thus did not contain inhibitors of BSF-1 activity.(ABSTRACT TRUNCATED AT 400 WORDS)     

Evolutionary aspects of trypanosomes: Analysis of genes

Summary The genes for four glycolytic enzymes ofTrypanosoma brucei have been analyzed. The proteins encoded by these genes show 38–57% identity with their counterparts in other organisms, whether pro- or eukaryotic. These data are consistent with a phylogenetic tree in which trypanosomes diverged very early from the main branch of the eukaryotic lineage. No definite conclusion can be drawn yet about the evolutionary origin of glycosomes, the microbodies of trypanosomes which contain most enzymes of the glycolytic pathway. A bias could be observed in the codon usage of the glycolytic genes and genes for other housekeeping proteins, indicating that trypanosomes may have selected a nucleotide sequence that enables efficient translation. However, the genes for variant surface glycoproteins (VSGs) do not show such a bias. This lack of preference for special codons is explained by the high evolutionary rate that could be observed for VSG genes.Presented at the FEBS Symposium on Genome Organization and Evolution, held in Crete, Greece, September 1–5, 1986