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1.
Guanylate cyclase from crude homogenates of vegetative has been characterized. It has a pH optimum of 8.0, temperature optimum of 25°C and requires 1 mM dithiothreitol for optimal activity. It strongly prefers Mn++ to Mg++ as divalent cation, requires Mn++ in excess of GTP for detectable activity, and is inhibited by high Mn++ concentrations. It has an apparent Km for GTP of approximately 517 μM at 1 mM excess Mn++.The specific activity of guanylate cyclase in vegetative homogenates is 50–80 pmoles cGMP formed/min/mg protein. Most of the vegetative activity is found in the supernatant of a 100,000 x g spin (S100). The enzyme is relatively unstable. It loses 40% of its activity after 3 hours storage on ice. Enzyme activity was measured from cells that had been shaken in phosphate buffer for various times. It was found that the specific activity changed little for at least 8 hours. Cyclic AMP at 10?4 M did not affect the guanylate cyclase activity from crude homogenates of vegetative or 6 hour phosphate-shaken cells. 相似文献
2.
Evelyn A. Devine Mary C. Moran Peter J. Jederlinic Anthony J. Mazaitis Henry J. Vogel 《Biochemical and biophysical research communications》1975,67(4):1589-1593
The transducing phage λd, carrying a portion of the chromosome including , is derived from the heat-inducible, lysis-defective strain λy199, which has the and deletions. Cleavage of λy199 DNA by RI endonuclease, followed by agarose slab gel electrophoresis, results in bands corresponding to the known C, D, E, and F segments of λ, and a segment A′ (A plus B minus minus , the cleavage site between A and B being eliminated). Cleavage of λd DNA by RI yields the expected D, E, and F segments of λ and four other segments, termed 14-1 through 14-4, whose length is 17.5, 6.2, 3.0, and 2.0 kilobases, respectively, as determined by electron microscopy and corroborated by electrophoretic mobility. Heteroduplex analysis shows that the cluster is on the 14-1 segment. 相似文献
3.
Ronald M. Hamelik Mead M. McCabe 《Biochemical and biophysical research communications》1982,106(3):875-880
An inhibitor of , endodextranase was detected in proteins prepared from batch cultures of , strains representing serotypes through . Affinity chromatography of strain 6715-49 proteins, which apparently were free of endodextranase activity, yielded an active endodextranase and, in a separate peak, the endodextranase inhibitor. The presence of the inhibitor in culture fluids accounts for the absence of endodextranase activity in batch-grown cultures of , known to produce this enzyme. 相似文献
4.
Yuji Kamiya Akira Sakurai Nobutaka Takahashi 《Biochemical and biophysical research communications》1980,94(3):855-860
Rhodotorucine which induces mating tube formation of cells in is metabolized rapidly by cells. By use of labeled rhodotorucine , the degradation was found to be proteolytic. Two peptide fragments Tyr-Pro-Glu-Ile-Ser-Trp-Thr-Arg and Asn-Gly-Cys(S-farnesyl) were identified as the metabolites. Proteolysis of the pheromone mainly occurred on the cell surface. Culture filtrate of cells at log phase did not metabolize rhodotorucine . 相似文献
5.
J.A. Pérez-González A. Jiménez 《Biochemical and biophysical research communications》1984,125(3):895-901
The paromomycin producing organism is resistant to this antibiotic and contains a phosphotransferase which inactivates paromomycin. The gene encoding this enzyme has been inserted in the vector pIJ702 and then cloned in , selecting for paromomycin-resistance. Three plasmids have been isolated and one of them, pMJ1, contains a 2.2 kb insert with a single HindIII restriction site. Insertion of foreign DNA in this site blocks the expression of the phosphotransferase enzyme indicating that it is within the cloned gene. These findings provide a new dominant selective marker for cloning vectors with the versatility of insertional inactivation. 相似文献
6.
OKY-1581 is an effective inhibitor of thromboxane synthesis and . The generation of thromboxane B2 (TxB2), prostaglandin E (PGE) and prostaglandin F (PGF) was measured following clotting and during platelet aggregation induced by collagen. The presence of OKY 1581 either or caused a reduction in TxB2 generation during clotting and platelet aggregation with a concomitant increase in PGE and PGF. The effect could be observed two hours after oral or subcutaneous administration of 5 to 100 mg per rabbit and lasted for 24 to 48 hours. The reduction in TxB2 was not accompanied by an inhibition of clotting or platelet aggregation. OKY-1581 appears to be a suitable agent for studying the role of TxB2 in atherosclerosis. 相似文献
7.
Robert C. Richmond John D. Zimbrick 《Biochemical and biophysical research communications》1975,64(1):391-398
Cells of C thy?321 are examined for thymine residue release from DNA following gamma-irradiation from 5 to 15 krad. Experimental conditions are designed to inhibit enzyme activity that might promote base residue release. Enzyme action is restricted in order to assess the physicochemical action of radiation on cellular DNA, and to this end irradiation is done under O2, N2, and N2O saturating conditions. Both thymine and thymidine release from bacterial DNA are detected and quantitated, and three oxygen effects are noted in comparing yields of these products. No difference in effect is observed between N2 and N2O gassing conditions, suggesting that the hydroxyl radical has little effect on thymine or thymidine release from irradiated DNA . 相似文献
8.
Josie M. Nichols G.A.M. Cross 《Biochemical and biophysical research communications》1976,71(3):796-802
Kinetoplast DNA from the order Kinetoplastidae (trypanosomatids) exists as large associations (molecular weight 4 × 1010), made up of about 104 small, probably circular, molecules, commonly known as ‘minicircles’. These minicircles were originally thought to be identical in base composition, suggesting that the coding capacity of kinetoplast DNA is very restricted. However, linear molecules have also been observed in preparations of kinetoplast DNA, which, if they contain unique sequences, could represent additional genetic information. This linear DNA has been assumed to be derived from the kinetoplast, but the possibility of it being nuclear contamination has not been definitely ruled out. Work presented in this paper demonstrates that nuclear DNA contamination may indeed be present in kinetoplast DNA prepared by a commonly used method. 相似文献
9.
G. Sheir-Neiss R.V. Nardi M.A. Gealt N.R. Morris 《Biochemical and biophysical research communications》1976,69(2):285-290
[35S] labeled extracts of the fungus were copolymerized with purified porcine brain tubulin. The [35S] protein which copurified with porcine microtubules was found to be similar to [3H] chick tubulin when the two were coelectrophoresed on several polyacrylamide gel electrophoresis systems. These results strongly suggest the presence in of a tubulin-like protein. 相似文献
10.
This case report addresses the occurrence of Brucellosis and its effect on the cattle in developing countries. Three Zebu bulls () are presented and the clinical and pathologic signs are described. Conception rates declined following an abortion storm in one herd and without prior abortions in another herd. Semen collected by electro-ejaculation was found to be azoospermic or with very few spermatozoa. was isolated from seminal vesicles, testes and epididymides. Organs affected and showing microscopic lesions were testes, epididymides and seminal vesicles. The latter were not consistently affected. None of the bulls showed impairment of libido or breeding capacity. 相似文献
11.
The specific synthesis of F mRNA directed by the F gene carried on the specialized transducing bacteriophage F, performed , is described with the use of an S180 extract from a strain carrying R?. Synthesis of F mRNA is biphasic at approximately 7 minutes. The regulation of F mRNA synthesis by the specific arginine holorepressor present in an S180 extract prepared from a strain carrying the R+ allele is described. 相似文献
12.
Uterine stage embryos collected from the hamster (8-cell) and cow (morula, early blastocyst) were monitored for development (embryo culture) and (embryo transfer) following premature removal of the zona pellucida.Removal of the zona pellucida did not significantly affect development to the blastocyst stage of (1) 8-cell hamster embryos (zonae removed by a combined enzymic-mechanical procedure), (2) bovine morulae (zonae removed by mechanical means only) (3) early bovine blastocysts (zonae removed by the enzymic-mechanical technique).Zona-free hamster embryos formed significantly fewer viable fetuses than did zona-intact embryos. The lower incidence of fetal development observed following transfer of zona-free 8-cell hamster embryos may have resulted in part from the formation of chimeras by fusion of these embryos . Such fusion was observed to occur between zona-free embryos placed in close proximity. The proportion of pregnancies resulting from transfer of bovine blastocysts cultured from zona-free morulae was similar to that of zona-intact embryos.In this study we have demonstrated that (1) enzymic and mechanical procedures used to remove zonae pellucidae from uterine-stage hamster and bovine embryos do not adversely affect subsequent development of these embryos and and (2) zonae pellucidae are not required for normal development of these embryos. These findings have implications for microsurgery of mammalian embryos and for embryo transfer. 相似文献
13.
In an attempt to isolate and to study the electron transport system of , we have isolated and purified a membrane-bound cytochrome . The cytochrome , purified as a detergent (Triton X-100) and hemoprotein complex, contained 1.6 nmoles heme per mg of protein. Cold-temperature spectrum showed that no other cytochrome was associated with the purified preparation, and electrophoresis revealed that only one type of hemoprotein was obtained. The purified cytochrome reacted with both carbon monoxide and cyanide readily. Only in the reduced form did it combine with carbon monoxide, whereas the oxidized form reacted with cyanide. An “oxygenated” form of the cytochrome was demonstrated to be spectrally distinguishable from both the oxidized and the reduced forms. 相似文献
14.
The synthesis and characterization of - and -3,4-bis(4-hydroxyphenyl)-2-hexene (- and -pseudo-DES) and of -3,4-bis(4-hydroxyphenyl)-2-hexen-1-ol (-1-hydroxypseudo-DES) are described. These compounds are useful as probes in the study of hormone action. 相似文献
15.
16.
Yolk free blastoderms of chick embryo were incubated 3 or 22 hours with labeled pregnenolone, progesterone, 17-hydroxyprogesterone, dehydro-epiandrosterone, androstenedione, testosterone and estradiol-17β. Metabolites and unconverted substrates were found both in the incubation medium and in the cells. Enzymes responsible for identified conversions were: 17α-hydroxylase, 17-20-desmolase, Δ53β- and 3α-hydroxysteroid dehydrogenase, 17β-hydroxysteroid dehydrogenase and 5α- and 5β-reductase. The results suggest that the steroid metabolizing enzyme activities found may reflect a more general ability of early embryonic cells. 相似文献
17.
R. Rodriguez C. Lopez-Otin D. Barber J.L. Fernandez-Luna G. Gonzalez E. Mendez 《Biochemical and biophysical research communications》1982,108(1):315-321
The NH2-terminal amino acid sequence of the three anti-tumor proteins, alfa-sarcin, mitogillin and restrictocine, has been determined for 20 cycles by automated sequencing procedure. A high degree of sequence homology was observed in this region of the molecule. In addition, extensive sequence homology, ranging from 65 to 100% was found in three other carboxymethylcysteine-containing peptides isolated and sequenced from each molecule. 相似文献
18.
James B. Howard Thomas Lorsbach Lawrence Que 《Biochemical and biophysical research communications》1976,70(2):582-588
In 80% dimethyl sulfoxide/H2O, ferredoxin FeS clusters can be extruded with benzene thiol. The extruded clusters have an absorption spectra maximum at 458 nm which is characteristic of 4Fe4S centers. The amino terminal sequence of the ferredoxin has 7 of the 8 Cys residues at residue numbers 8, 11, 16, 20, 24, 39 and 42. Except for Cys 24, all of these residues can be correlated to homologous Cys residues in other bacterial ferredoxins. Although two thirds of the first 45 residues are identical to or conservative replacements for the first 43 residues of other bacterial ferredoxins, the insertion of Cys-24 indicates a major change in the environment of one of the two 4Fe4S clusters. 相似文献
19.
Daniel H. Riddick Anthony A. Luciano William F. Kusmik Ila A. Maslar 《Life sciences》1978,23(19):1913-1921
Relatively large amounts of immunoreactive prolactin were measured in homogenates of human decidual tissue obtained immediately after delivery of normal term pregnancies. In order to study the release and possible synthesis of prolactin by this tissue, explants of decidua were incubated for 24 hours at 37°C in oxygenated Gey's buffer containing 20% fetal calf serum. When cycloheximide was added to the medium in concentrations sufficient to prevent protein synthesis, 85–90% of the prolactin present in the tissue was released into the medium during the first 3 hours of incubation. No additional prolactin accumulated in either the medium or the tissue during the remainder of the incubation period. In the absence of cycloheximide, the prolactin concentration in the medium increased progressively during incubation, so that after 24 hours the total amount of hormone present in the tissue and medium was significantly greater than that in the tissue and medium prior to incubation (37.6 ± 9.6 ng/ml at 0 time vs 82.2 ± 7.7 ng/ml at 24 hours). When 3H-1-leucine (100 u Ci) was supplied during incubation, radioactive proteins were detected in the medium at 24 hr, 14–20% of which were specifically precipitated by antiserum to human pituitary prolactin. When aliquots of this medium were chromatographed on Sephadex G-100, 80–95% of the 3H-proteins precipitated by antiserum to pituitary prolactin eluted in the same position as did purified, iodinated pituitary prolactin. These data indicate that a species of prolactin which is identical to pituitary prolactin by the criteria of immunoprecipitation and gel chromatography is synthesized by human decidual tissue . 相似文献
20.
Robert D. Koos Martin R. Clark Per O. Janson Kurt E.B. Ahrén William J. LeMaire 《Prostaglandins & other lipid mediators》1983,25(5):715-724
Prostaglandin (PG) levels in follicular fluid from preovulatory follicles of rabbit ovaries perfused were measured in order to compare PG changes in this model system with those that occur and in isolated, LH-treated follicles . One ovary from each rabbit was perfused without further treatment (control). The other ovary was exposed to LH (0.1 or 1 ug/ml) beginning 1 hour (h) after initiation of perfusion. Samples of perfusion medium were taken at frequent intervals for measurement of PGE, PGF, progesterone and estradiol 17β. The perfusions were terminated when the first ovulation occurred or appeared imminent as judged by changes in the size and shape of the follicles. Follicular fluid was then rapidly aspirated from all large follicles on both ovaries for PGE and PGF measurement.Ovulations occurred only in the LH-treated ovaries. Progesterone and estradiol levels were significantly elevated in the perfusion medium within 1 h of LH treatment in comparison to controls. PG levels in perfusion medium from the control and LH-treated ovaries were not different throughout perfusion and increased in both groups. In contrast, PG levels measured in follicular fluid from LH-treated ovaries were 4- to 5-fold greater than in fluid from control ovaries. It is concluded that ovulation induced by LH in this experimental model is accompanied by an increase in follicular PG levels similar to that seen in other and models. This difference in follicular PG levels between the LH-treated and control ovaries is, however, not reflected in the perfusion medium. 相似文献