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91.
Isolation and characterization of nucleases from a clinical isolate of Serratia marcescens kums 3958 总被引:1,自引:0,他引:1
Two new extracellular nucleases, nucleases SM1 and SM2, were purified from the culture fluid of S. marcescens kums 3958, a fresh clinical isolate. The purification was carried out by the following steps; ammonium sulfate precipitation, and DEAE-cellulose and Sephadex G-100 column chromatography. At the final step, nucleases SM1 and SM2 were purified about 3,700- and 1,000-fold, respectively. They were free from phosphomonoesterase and phosphodiesterase activities. The pIs were 8.1 and 7.5 for nucleases SM1 and SM2, respectively. The molecular weight was estimated to be 35,000 for both enzymes by SDS-polyacrylamide disc gel electrophoresis. The results of amino acid analyses showed that both the threonine and serine contents were higher in nuclease SM2 than in SM1. Furthermore, nuclease SM1 was more stable than nuclease SM2 at 4 degrees C. The other properties of the two enzymes were similar; pH optimum (8.0), Mg2+ or Mn2+ for activation, and inhibition by chemical reagents such as EDTA and pyrophosphate. No significant difference was found in base specificity between nucleases SM1 and SM2. Both enzymes specifically degraded double-stranded homopolymers, especially poly(I). poly(C), as well as yeast RNA and calf thymus DNA. They hardly degraded, however, single-stranded homopolymers such as poly(dA), poly(G), and poly(U). 相似文献
92.
Isolation and characterization of a third proteoglycan (PG-Lt) from chick embryo cartilage which contains disulfide-bonded collagenous polypeptide 总被引:11,自引:0,他引:11
A Noro K Kimata Y Oike T Shinomura N Maeda S Yano N Takahashi S Suzuki 《The Journal of biological chemistry》1983,258(15):9323-9331
Chick embryo epiphyseal cartilage has been shown to contain three different proteoglycan species (PG-H, PG-Lb, and PG-Lt). This report is concerned with the purification and characterization of the third proteoglycan, PG-Lt. The proteoglycan can be separated from the other two by virtue of its low buoyant density in a CsCl density gradient and further purified by consecutive ion exchange and gel chromatography. The final preparation is composed of PG-Lt monomer and PG-Lt oligomer. The amino acid composition of PG-Lt is quite different from that of PG-H and PG-Lb and rather resembles that of collagens with respect to high content of glycine and high degrees of hydroxylation of proline and lysine. PG-Lt monomer is composed of disulfide-bonded subunits of Mr congruent to 120,000 and 190,000 as demonstrated by its gel electrophoretic behavior after reduction with 2-mercaptoethanol. The latter, but not the former, contains dermatan sulfate chains with glucuronic acid/iduronic acid residues and yields a protein-enriched core molecule of Mr congruent to 100,000 after digestion with chondroitinase ABC. Both of the protein subunits are completely digestible with bacterial collagenase. Immunofluorescence microscopic examination of cartilage tissues, using an antibody against PG-Lt, shows that this proteoglycan exists in both the cartilage matrix and perichondrial noncartilagenous region. When chondrocytes are plated onto tissue culture dishes, the antibody stains strands found on the cell surfaces and in the intercellular space of substrate-attached cell layers, suggesting that PG-Lt mediates cell-to-cell and cell-to-substrate contacts. 相似文献
93.
Makoto Hayashi Kazuko Fukushima Akihide Kitamura 《Chemistry and physics of lipids》1983,33(3):233-239
Krafft points of diacylglycerophosphocholines (PC) were measured in alkanes-cyclohexane solutions by differential scanning calorimetry, and it was found that they were regularly increased following the increase in alkane content in the solutions and the chain length of the alkanes. From these results it was deduced that the mixing of PC with alkanes occurred in the gel state of the PC, but not in micelles at higher temperatures above the Krafft points. where micellar solutions are provided. The penetration of alkanes into gel state PC was found to be dominated by Langmuir type interaction, and the affinity of alkanes increases with increasing in chain lengths. Above the Krafft points, the micelle formation was confirmed by using the fluorescence probe technique. 相似文献
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97.
Makoto Nagoshi 《Population Ecology》1966,8(1):20-36
- In order to get the knowledge on the age composition of “isaza” population in Lake Biwa and the effect of population density on growth, monthly distribution of mean body length and mean body weight has been analyzed on the basis of monthly haul by “isazabiki” trawl during 1949 to 1953 and also 1960 to 1965.
- There is no apparent sex difference in the growth in the first and second years of life.
- “Isaza” population is composed of two age groups, age 0 and 1 groups (1+fish), the latter occupying by far the greater part in commercial catch.
- During the growth season fishes of both ages feed mainly on zooplankton, though in winter frequently take chironomid larvae, gammaridae and others in volume.
- The growth season falls in the period from April to October in both age groups.
- A considerable yearly variation occurring in growth is in close connection with the fluctuation of population density of all ages.
- The influence affected by the density of age 1 group is larger than that by age 0 group.
98.
Makoto Endo 《Ichthyological Research》1989,36(3):346-349
Glomerular vasculature was investigated in the carpCyprinus carpio, the scorpionfishSebastiscus marmoratus, and the marine catfishPlotosus lineatus with the resin-replica method. An afferent arteriole was connected with a glomerulus in every fish. It was slender in the
carp, whereas the scorpionfish and marine catfish possessed thick afferent arterioles. The glomerular capillaries were sinusoidal.
The divergences, convergences, and windings of these capillaries were not well developed in any of the fish. The glomerular
capillaries converged into an efferent arteriole in the carp and scorpionfish. In the marine catfish, on the other hand, most
of the glomeruli had two efferent arterioles. 相似文献
99.
Makoto Suematsu Shigenari Houzawa Soichiro Miura Hiroshi Nagata Tetsuji Kitahora Tetsuo Morishita Chikara Oshio Masaharu Tsuchiya 《Luminescence》1989,4(1):531-534
Reaction difference of oxyradical generation and luminol-dependent photoemission of zymosan- and phorbol ester-treated neutrophils were investigated using a conventional photomultiplier and ultrasensitive photonic imaging technique. Zymosan-treated cells released a concentrated photonic burst corresponding to the cellular distribution. In contrast, phorbol ester-treated cells produced a negligible level of photoemission, and the additional application of Ca2+ ionophore enhanced the photonic burst, which was gradually spread out into extracellular space. Serine protease inhibitors did not attenuate PMA-induced chemiluminescence but did attenuate zymosan-induced chemiluminescence. This suggests the involvement of serine protease in the respiratory burst of phagocytizing neutrophils. 相似文献
100.
A highly sensitive chemiluminescent assay for NAD(P)H have been developed. The principle of the method is as follows; NAD(P)H reduces molecular oxygen to superoxide anion (O) and hydrogen peroxide (H2O2) in the presence of 1-methoxy-5-methylphenazinium methyl sulphate (1-MPMS) as electron mediator. The produced O and H2O2 can be measured by chemiluminescent reaction using isoluminol (IL) and microperoxidase (m-POD). A linear relationship between chemiluminescence intensity and NAD(P)H concentration (log/log) was obtained ranged from 10?9 mol/I to 10?5 mol/I. This chemiluminescent reaction has been coupled to the assay of glucose-6-phosphate dehydrogenase (G6PDH), β-D -galactosidase (β-Gal) and alkaline phosphatase (ALP). The detection limits of G6PDH, β-Gal and ALP were 10?18 mol, 10?20 mol and 10?18 mol per assay, respectively. The chemiluminescent assay of these enzymes applied to chemiluminescent enzyme immunoassay for 17α-hydroxy-progesterone and DNA hybridization assay using these enzymes as label. 相似文献