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1.
Purification of a soluble, sodium-nitroprusside-stimulated guanylate cyclase from bovine lung 总被引:7,自引:0,他引:7
A soluble, sodium-nitroprusside-stimulated guanylate cyclase as been purified from bovine lung by DEAE-cellulose chromatography, ammonium sulfate precipitation, chromatography on Blue Sepharose CL-6B and preparative gel electrophoresis. Apparent homogeneity was obtained after at least 7000-fold purification with a yield of 3%. A single stained band (Mr 72000) was observed after gel electrophoresis in the presence of sodium dodecyl sulfate. The purified enzyme migrated as one band also under non-denaturing conditions in acrylamide gels (5-12%). The mobility of this band corresponded to an Mr of 145000. The enzyme sedimented on sucrose gradients with an S20, w of 7.0 S. Gel filtration yielded a Stokes' radius of 4.6 nm. These data suggest that the enzyme has an Mr of approximately 150000 and consists of two, presumably identical, subunits of Mr 72000. Sodium nitroprusside stimulated the purified enzyme 15-fold and 140-fold to specific activities of 8.5 and 15.7 mumol of cGMP formed min-1 mg-1 in the presence of Mn2+ and Mg2+, respectively. Formation of cGMP was proportional to the incubation time and to the amount of enzyme added. The stimulatory effect of sodium nitroprusside was half-maximal at about 2 microM, was observed immediately after addition and could be reversed either by dilution or by removal of sodium nitroprusside on a Sephadex G-25 column. The purified enzyme in the absence of catalase was stimulated by sodium nitroprusside, N-methyl-N'-nitro-N-nitrosoguanidine and 3-morpholino-sydnonimine and in the presence of catalase by sodium nitrite and sodium azide. In the presence of Mn2+ and sodium nitroprusside, the purified enzyme catalyzed the formation of cAMP from ATP at a rate of 0.6 mumol min-1 mg-1. 相似文献
2.
B Hofmann E Langhoff B O Lindhardt N Odum J J Hyldig-Nielsen L P Ryder P Platz B K Jakobsen K Bendtzen N Jacobsen 《Cellular immunology》1989,121(2):336-348
Retroviral infections are accompanied by immunosuppression in a variety of species. For feline leukemia virus, the immunosuppression has been ascribed to the transmembrane envelope protein, p15E, which suppresses the proliferative responses of cat, mouse, and human lymphocytes. A similar suppressive effect has been shown for a lysate of human immunodeficiency virus (HIV), strain HTLV-IIIB. Here we determined that detergent-disrupted HTLV-IIIB lystate exerted a strong suppressive effect on PHA-stimulated lymphocytes. Preparations of whole virions, a lysate of a local HIV isolate grown on MP-6 cells, and a commercially obtained UV and psoralene-inactivated lysate were examined and demonstrated to have a similar suppressive effect. The HIV lysate was not directly cytotoxic to lymphocytes and did not contain tumor necrosis factor or lymphotoxin. The HIV lysate specifically suppressed the proliferation of a range of hemopoietic cell lines from man and mouse including three EBV transformed CD4- and IL-2 receptor-negative B-cell lines. The lysate also suppressed the formation of human bone marrow colonies, whereas the lysate had only a slight or no effect on fibroblasts. The suppression of lymphocyte proliferation was not abrogated by addition of IL-2 or IL-1 and the HIV lysate inhibited the expression of IL-2 receptors on suboptimal PHA-stimulated mononuclear cells. The suppressive factor(s) has not been characterized in molecular terms, but suppressive activity was recovered in fractions with a molecular weight of about 67,000 and in both the glycoprotein fraction and in the glycoprotein-depleted fraction of the HIV lysate. Sera from one-third of a small series (N = 13) of individuals with antibodies to HIV seem to be able to neutralize the suppressive properties of HIV lysate in cultures. 相似文献
3.
The complete primary structure of the alpha- and beta-chains of the hemoglobin of Malayan Sun Bear (Helarctos malayanus) is presented. After cleavage of the heme-protein link and chain separation by RP-HPLC, amino-acid sequences were determined by Edman degradation in liquid- and gas-phase sequenators. An interesting result of this work is the demonstration that the hemoglobin of Malayan Sun Bear is identical to the hemoglobins of Polar Bear (Ursus maritimus) and Asiatic Black Bear (Ursus tibetanus). The paper gives an updated table of identical hemoglobin chains from different species. This paper may be considered as a compilation of work on the genetic relationship of Pandas. 相似文献
4.
Cladocera in space and time: Analysis of lake sediments 总被引:6,自引:6,他引:0
Wolfgang Hofmann 《Hydrobiologia》1987,145(1):315-321
Shells of Bosminidae and Chydoridae are quantitatively preserved in lake sediments. The chronological deposition of these
remains provides the means for longterm observation of these Cladocera, both in terms of species and communities.
Chydorid analysis, as based on subfossil assemblages, is an analysis of community and provides direct observation of community
dynamics over extended periods of time. It has proved to be a valuable method to obtain information on the influence of environmental
factors and time on community characteristics.
Morphological variation inBosmina (Eubosmina) has been followed for some thousand years. This is of special interest for the evaluation of taxonomic rank (species, forms)
if closely related taxa have co-existed.
Bosmina successions, as well as shifts in the chydorid fauna, are related to environmental change. Thus, cladoceran analysis of lake
sediments provides information on the developmental history of lakes and allows observation of the effects of longterm environmental
changes, such as climatic changes and eutrophication. 相似文献
5.
6.
Evolutionary steps of ecophysiological adaptation and diversification of ruminants: a comparative view of their digestive system 总被引:23,自引:0,他引:23
R. R. Hofmann 《Oecologia》1989,78(4):443-457
Summary A review is made of the ruminant digestive system in its morphophysiological variations and adaptations relating to foraging behaviour, digestive physiology, to interactions between plants and ruminants and to geographic and climatic diversity of ruminants' ecological niches. Evidence is provided for evolutionary trends from an extreme selectivity mainly for plant cell contents and dependence upon a fractionated fore- and hindgut fermentation, to an unselective intake of bulk roughage subjected to an efficient plant cell wall fermentation, mainly in the forestomachs. The review is based on detailed comparative morphological studies of all portions of the digestive system of 65 ruminant species from four continents. Their results are related to physiological evidence and to the classification of all extant ruminants into a flexible system of three overlapping morphophysiological feeding types: concentrate selectors (40%), grass and roughage eaters (25%) and intermediate, opportunistic, mixed feeders (35%). Several examples are discussed how ruminants of different feeding types are gaining ecological advantage and it is concluded that ruminants have achieved high levels of digestive efficiency at each evolutionary stage, (including well-documented seasonal adaptations of the digestive system) and that ruminant evolution is still going on. Deductions made from the few domesticated ruminant species may have, in the past, biased scientific evaluation of the free-ranging species' ecology. The main threat to a continuous ruminant evolution and diversity appears to be man's neglect for essential ecological interactions between wild ruminants and their specific habitats, which he alters or destroys.Abbreviations
bw
body weight
-
CS
concentrate selector
-
DFC
distal fermentation chamber (distended caecocolon)
-
GR
grass and roughage eater
-
IM
intermediate (mixed) feeder
-
PFC
proximal fermentation chamber (ruminoreticulum/forestomachs)
-
RR
Ruminoreticulum
-
SCFA
Short-chain fatty acis (acetic, butyric, propionic acid set free by rumen bacteriae)
-
SE
Surface enlargement (of absorptive mucosa)
Supported by German Research Community grant DFG Ho 273/6Dedicated to Professor Dr. Dr. h.c. Dietrich Starck on the occasion of his 80th birthday 相似文献
7.
Ultrastructural localization of WGA,RCA I,LFA and SBA binding sites in the seven-day-old mouse embryo 总被引:1,自引:0,他引:1
Summary Transglutaminases are Ca2+-dependent intra-and extracellular enzymes catalyzing the cross-linking between proteins and/or polyamines, thereby eliciting
divergent physiological effects such as fibrin clot stabilization or hair follicle cross-linking. A secretory transglutaminase
(EC 2.3.2.13) was isolated from the coagulating gland of the rat. The protein is highly glycosylated. A fraction purified
to homogeneity was used as an antigen to raise polyclonal antibodies in rabbits. These antibodies were used to identify the
secretion sites of the protein within the male accessory sex glands as well as to study the immunological relationships of
the respective antigen within different organs of different species. In the rat, the coagulating gland and likewise the dorsal
prostate gave a positive immunoreaction. In the guinea pig, a closely related protein was detected in the anterior prostate.
No cross-reactivity was found with membrane-bound transglutaminase from liver, erythrocytes or blood clotting factor XIIIa.
The intraluminal secretion of the aforementioned glands was only weakly stained. No secretory granules were observed in the
glandular epithelium but instead bleb-like structures reminiscent of apocrine secretion. A slight background stain of the
epithelium remained even in castrated animals where secretion is largely suppressed. The background stain is attributed to
a tissue-type, membrane-bound, non-secretory transglutaminase that is not androgen dependent, but instead synthesized only
after androgen deprivation. 相似文献
8.
H P Zassenhaus T J Hofmann R Uthayashanker R D Vincent M Zona 《Nucleic acids research》1988,16(8):3283-3296
The nuclear gene from Saccharomyces cerevisiae that encodes the major mitochondrial nuclease was cloned. Gene sequences were identified from a lambda gt11 library by antibodies specific to the mitochondrial nuclease. DNA from the phage recombinant was used to isolate the entire nuclease gene from a plasmid library. Yeast strains containing the nuclease gene on a multicopy plasmid vector overproduced mitochondrial nuclease 20-40 times relative to a wild-type strain. Strains containing a null allele of the nuclease gene lacked all traces of mitochondrial nuclease. Both cell types, however, were phenotypically wild-type indicating that the nuclease is not an essential enzyme for mitochondrial function. The locus encoding the mitochondrial nuclease is termed NUC1. 相似文献
9.
10.
An efficient procedure for obtaining nor-bile acids from natural (C24) bile acids is described. Treatment of formylated bile acids with sodium nitrite in a mixture of trifluoroacetic anhydride with trifluoroacetic acid gives, through a "second order" Beckmann rearrangement, 24-nor-23-nitriles. These compounds, on alkaline hydrolysis, afford the corresponding nor-bile acids in high yields. The sequence was successfully applied to the synthesis of 3 alpha-hydroxy-24-nor-5 beta-cholan-23-oic (norlithocholic) acid, 3 alpha,6 alpha- (norhyodeoxycholic), 3 alpha,7 alpha- (norchenodeoxycholic), 3 alpha,7 beta- (norursodeoxycholic), and 3 alpha,12 alpha-dihydroxy-24-nor-5 beta-cholan-23-oic (nordeoxycholic) acids, as well as 3 alpha,7 alpha,12 alpha-trihydroxy-24-nor-5 beta-cholan-23-oic (norcholic) acid. 13C-NMR spectra of their methyl esters are reported. The procedure provides a more rapid alternative to the Barbier-Wieland degradation for shortening by one methylene group the side chain of natural (C24) bile acids. 相似文献