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1.
Tissue distribution of beta 1- and beta 2-subunits of regulatory guanine nucleotide-binding proteins 总被引:3,自引:0,他引:3
K D Hinsch I Tychowiecka H Gausepohl R Frank W Rosenthal G Schultz 《Biochimica et biophysica acta》1989,1013(1):60-67
The beta-subunit of G-proteins occurs in two forms (beta 1 and beta 2), which differ in their primary structure as derived from cDNA clones and in their mobilities on SDS gels (36 and 35 kDa, respectively). To assess the tissue distribution of the two forms of beta-subunits, we synthesized peptides corresponding to defined regions of beta 1- and beta 2-subunits and injected them into rabbits; the antisera obtained reacted either with both beta-subunits or specifically with the beta 1- or the beta 2-subunit. They were used to identify the two beta-subunits in membranes prepared from various rat tissues and from human placenta. The concentration of total beta-subunits was high in rat brain and lung, human placenta, rat kidney, liver and spleen; it was much lower in rat erythrocytes, cardiac and skeletal muscle. In all tissues studied, both beta 1- and beta 2-subunits were detectable. In most tested tissues, the two forms were about equally distributed, whereas in the placenta, the beta 2-subunit was found to occur in approx. 2-fold excess over the beta 1-subunit. Our results demonstrate that both beta-subunits are widely distributed. In the majority of tissues, levels of beta 2-subunits are very similar to those of beta 1-subunits. Thus, the abundance of beta 2-subunits as compared to that of the beta 1-subunit is considerably higher than was previously estimated by measuring the respective mRNA levels. 相似文献
2.
Cyclic AMP-independent, dual regulation of voltage-dependent Ca2+ currents by LHRH and somatostatin in a pituitary cell line. 总被引:6,自引:1,他引:5
W Rosenthal J Hescheler K D Hinsch K Spicher W Trautwein G Schultz 《The EMBO journal》1988,7(6):1627-1633
Voltage-dependent Ca2+ currents appear to be involved in the actions of hormones that regulate pituitary secretion. In order to investigate modulation of Ca2+ currents by release-inducing and release-inhibiting hormones, we performed whole-cell clamp experiments in the pituitary cell line GH3. The resting potential was approximately -40 mV; spontaneous action potentials were observed in the majority of cells. Superfusion of cells with the stimulatory hormone, LHRH, depolarized the plasma membrane to approximately -10 mV, whereas the inhibitory hormone, somatostatin, caused hyperpolarization to approximately -60 mV; both hormones suppressed spontaneous action potentials. Under voltage clamp conditions, GH3 cells exhibited slowly and fast inactivating Ca2+ currents. LHRH increased whereas somatostatin decreased the slowly inactivating currents; fast inactivating currents were not affected by these hormones. The stimulatory effect of LHRH was not mimicked by intracellularly applied cAMP. In contrast to vasoactive intestinal peptide and forskolin, LHRH did not activate adenylate cyclase in membranes of GH3 cells, but rather appeared to cause inhibition of the enzyme. Hormonal stimulation and inhibition of inward currents were abolished by pretreatment of the cells with pertussis toxin. In membranes of GH3 cells, we identified a pertussis toxin-sensitive G-protein of the Gi-type and Go. We conclude that LHRH and somatostatin modulate voltage-dependent Ca2+ currents via cAMP-independent mechanisms involving pertussis toxin-sensitive G-proteins. The occurrence of both pertussis toxin-sensitive hormonal stimulation and inhibition of voltage-dependent Ca2+ currents in one cell type suggest that these opposite regulations are mediated by distinct G-proteins. 相似文献
3.
Basement membranes were separated from rat glomeruli and purified by mild procedures, which led to a highly enriched basement membrane fraction. Here, the production and characterization of five monoclonal antibodies against tubular and glomerular basement membranes are described. These antibodies were analyzed immunohistochemically on frozen sections of rat, bovine, and human kidneys as well as on rat embryos. One monoclonal antibody (BM O II) exclusively recognized the glomerular basement membranes, another one (BM O VII) bound to tubular basement membranes and to Bowman's capsule. Three antibodies (BM O IV, BM M II, BM M III) recognized their antigens in both glomerular and tubular basement membranes as well as in mesangial cells. The BM O II antibody showed a stringent species specificity and bound only to glomerular basement membranes of the rat. The other four antibodies cross-reacted with human and bovine glomerular basement membrane and mesangial antigens; they also bound to other tissues in the developing rat embryo. Antibody binding to specific purified components of the basement membranes such as collagen type IV, laminin, heparan sulphate proteoglycan, and fibronectin was investigated by enzyme-linked immunosorbent assay (ELISA). None of these antibodies reacted with any of these known basement membrane components, indicating that the antibodies may serve as useful tools in future investigations of so far unidentified components of basement membranes. 相似文献
4.
Immunochemical detection of the alpha-subunit of the G-protein, GZ, in membranes and cytosols of mammalian cells 总被引:1,自引:0,他引:1
K Spicher K D Hinsch H Gausepohl R Frank W Rosenthal G Schultz 《Biochemical and biophysical research communications》1988,157(3):883-890
An antiserum (AS 98) was raised against a synthetic peptide deduced from published cDNA sequences of the alpha-subunit of the putative G-protein, GZ (Fong et al. Proc. Natl. Acad. Sci. USA 85, 3066-3070, 1988; Matsuoka et al. Proc. Natl. Acad. Sci. USA 85, 5384-5388, 1988). In membrane and cytosol preparations of many but not all tested mammalian tissues, AS 98 predominantly recognized two proteins of 40 and 43 kDa Mr. Whereas high levels of a 40 kDa GZ alpha-subunit were found in rat liver membranes and in brain cytosol, AS 98 failed to detect the alpha-subunit of GZ in brain membranes. 相似文献
5.
Sequence homologies of glucose-dehydrogenases of Bacillus megaterium and Bacillus subtilis 总被引:1,自引:0,他引:1
Peter Fortnagel Keith A. Lampel Klaus-Dieter Neitzke Ernst Freese 《Journal of theoretical biology》1986,120(4):489-497
The sequence homologies of the glucose dehydrogenase subunits of B. megaterium and B. subtilis are compared. From the known B. megaterium aminoacid sequence and the base sequence of the cloned B. subtilis structural gene we predict the B. megaterium structural glucose dehydrogenase gene. Assuming the minimal mutational changes to convert one gene into the other 23 transitions, 30 transversions, 1 inversion, 3 insertion-deletions, but no frameshifts are postulated necessary to interconvert the structural genes. The homology of both enzyme subunits of 85% reflects the close evolutionary distance between B. subtilis and B. megaterium. 相似文献
6.
Franz Bogner Michael Boppré Klaus-Dieter Ernst Jürgen Boeckh 《Journal of comparative physiology. A, Neuroethology, sensory, neural, and behavioral physiology》1986,158(6):741-749
Summary The tips of the labial palps ofRhodogastria possess a pit housing uniform sensilla (Fig. 1), histologically characterized by wall-pores and receptor cells with lamellated outer dendrites (Fig. 2). The receptor cell axons project to glomeruli in the deutocerebrum (cf. Fig. 3) which are not innervated by antennal receptors. From their histology as well as from their central projection these sense organs are identical with palpal pit organs of other Lepidoptera (Lee et al. 1985; Kent et al. 1986; Lee and Altner 1986).Physiologically, the palp-pit receptors respond uniformly; they are most excitable by stimulation with carbon dioxide (Fig. 6) while they exhibit relatively moderate responses to various odorants (Fig. 4). The responses to CO2 (Fig. 7) show a steep dose-response characteristic. In ambient atmosphere (i.e., ca. 0.03% CO2) the cells are in an excited condition already; the seeming spontaneous activity exhibited in air is decreased if the preparation is kept under N2 or O2 or CO2-free air (Figs. 7, 10). There is hardly any adaptation of the responses to continuous or repeated stimulation (Fig. 8). Perhaps CO2 sensitivity is correlated with sensilla characterized by both wall-pores and lamellated dendrites. Pilot tests indicate that CO2 perception might be widespread in the Lepidoptera (cf. Fig. 12), but the biological significance remains obscure. 相似文献
7.
Ultrastructural differentiation of the epithelium and mucous glands of the esophagus in the chick embryo 总被引:1,自引:0,他引:1
G W Hinsch 《Journal of morphology》1967,123(2):121-131
Esophagi of White Leghron chick embryos, six days to hatching, were fixed in 5% glutaraldehyde in a cacodylate-sucrose buffer, postfixed in 1% OsO4, dehydrated and embedded in Araldite. Tissues were sectioned with glass knives and viewed with a Philips EM 100C. The epithelium in early stages of development is characterized by small intercellular spaces, few cell processes and few desmosomes. In contrast the differentiated epithelium contains numerous cell processes, large intercellular spaces and numerous desmosomes. Mucin appeared in the mucous glands at 17 days. 相似文献
8.
9.
Structure of the female reproductive tract of the apple snail. II. Scanning electron microscopy 总被引:1,自引:0,他引:1
The albumen gland of Pomacea paludosa, a prosobranch gastropod, contains two main ducts. The albumen gland duct consists of a single layer of secretory and non-secretory cells. The surface of the non-secretory cells is covered with cilia. Microvilli are associated with the luminal edges of the secretory cells. Globules of secretory products appear at the cell surfaces. The capsule gland duct coils through the albumen gland and is composed of two opposing faces each of two layers of cells. The upper layer consists of ciliated non-secretory cells and the microvilli covered necks of the goblet-shaped secretory cells. The bases of the secretory cells comprises the lower layer of cells. Differences in the arrangement of cellular processes and number exists between the duct epithelia. 相似文献
10.
Paul Kuppert Siegrun Wilhelm Klaus-Dieter Spindler 《Journal of comparative physiology. B, Biochemical, systemic, and environmental physiology》1978,128(2):95-100
Summary From in vitro experiments using different binding assays it is in crayfish demonstrated that the cytosol of target tissues is able to bind both ecdysone and ecdysterone. The ability to bind ecdysteroids is destroyed by heating and by treatment with -chymotrypsin and N-ethyl-maleinimide (NEM) (Figs. 4, 5). In target tissues there is a strong positive correlation between protein content and binding (Fig. 6). The association of the hormone-protein-complex is rapid, taking only a few min even at 5° C (Fig. 3). The binding of the two hormones to the cytosol is both specific and saturable. The association constants for the cytoplasmic receptors from hypodermis, hindgut and midgut gland are in the range of 3–6×107 M–1 for ecdysone and 5–7×108 M–1 for ecdysterone (Fig. 8). The data suggest the existence of cytoplasmic ecdysteroid receptors. 相似文献