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991.
Characterization of calf liver glucosidase I and its inhibition by basic sugar analogs 总被引:4,自引:0,他引:4
Glucosidase I, the enzyme catalyzing the first step of N-linked oligosaccharide processing, has been purified from calf liver crude membranes [H. Hettkamp, G. Legler, and E. Bause, (1984) Eur. J. Biochem. 142, 85-90]. Binding experiments with concanavalin A-Sepharose suggest that glucosidase I is a glycoprotein with high-mannose carbohydrate chain(s). The enzyme has a subunit molecular mass of approximately 83 kDa and specifically hydrolyzes the terminal alpha-1,2-linked glucose residue from the natural Glc3-Man9-GlcNAc2 oligosaccharide. Studies with a variety of substrates modified in the aglycon moiety suggest that the Glc2 branch rather than the more distant domains of the substrate molecule are important for binding and hydrolysis. Glucosidase I does not require metal ions for activity and is strongly inhibited by 1-deoxynojirimycin (dNM) and its N-alkyl derivatives. Ki values range from 0.07 microM for N-methyl-dNM to 1.0 microM for dNM, measured at the pH-optimum of enzyme activity. The pH dependence of inhibition indicates that the cationic form of the inhibitors is the active species. Comparison of the Ki for N-decanoyl-dNM (approximately 70 microM) with that of N-decyl-dNM (approximately 0.4 microM) suggests that electrostatic interactions at the catalytic site of the enzyme are important for inhibitor binding. 1-Deoxymannojirimycin, previously assumed to be a specific mannosidase inhibitor, as well as its N-methyl and N-5-carboxypentyl derivatives, inhibit glucosidase I with Ki values around 190, 17, and 100 microM, respectively. This apparent lack of specificity shows that in vivo experiments on N-glycoprotein processing as well as the interpretation of results with these mannosidase inhibitors may give misleading results when these compounds are used in the millimolar range. 相似文献
992.
J J Candelier P Couillin P Eydoux A Boué 《Comptes rendus de l'Académie des sciences. Série III, Sciences de la vie》1986,302(8):303-308
The production of monoclonal antibodies against human embryonic renal cells allowed to display on the adult human kidney some antigens typical of certain structures or tissues: the proximal convoluted tubule for EG 9-11 and EG 19-6 monoclonal antibodies, the glomerular basement membrane for EG 14-1, the urothelium for EE 24-6, the connective tissue for EK 8-1 and EK 17-1 and probably the capsular and tubular basement membranes for EK 8-1. Simultaneously, we could follow the spatial and temporal repartition of the antigens during the renal development. One of them (EI 16-1) seemed to disappear in the adult and might correspond to a foetal type-antigen. 相似文献
993.
A recombination map of the human X-chromosome 总被引:2,自引:0,他引:2
Summary A family with 11 normal boys has been typed with DNA probes spanning the whole of the X-chromosome to observe directly the recombination events in 11 meioses from one female. This has (a) identified apparent recombination hot-spots on the X-chromosome; (b) shown the positions and numbers of cross-overs that have occurred in the p and q arms; (c) not shown any cross-overs in the centromeric region and (d) enabled the calculation of the genetic length of the X-chromosome. 相似文献
994.
995.
We show that expanded bed protein A affinity chromatography using Streamline rProtein A media is an efficient method for purifying a recombinant humanized monoclonal antibody from unclarified Chinese hamster ovary cell culture fluid and that it provides purification performance comparable to using a packed bed. We determined that the dynamic capacity of the expanded bed media is related to flow rate (measured in column volumes per hour) by a power function, which allows a high capacity at a low flow rate. At 250 cm h-1 with a 25 cm bed height (10 column volumes h-1), the dynamic capacity is 30 g l-1. The yield and purity (measured by the amount of host cell proteins, DNA, SDS-PAGE, and turbidity) of the antibody purified by expanded bed is comparable to the yield and purity obtained on a standard packed bed method using Prosep A media. 相似文献
996.
I. G. Minkevich A. D. Satroutdinov E. G. Dedyukhina T. I. Chistyakova E. N. Kaparullina A. V. Koshelev O. N. Okunev 《World journal of microbiology & biotechnology》2006,22(11):1205-1213
The effect of temperature on the maximum specific growth rate and the cell yield was studied during cultivation of two bacterial strains (LPM-4 and Pseudomonas sp. LPM-410) on EDTA under unlimited cell growth conditions in a pH-auxostat. Both strains displayed linear dependence of reciprocal biomass yield against reciprocal specific growth rate, from which the values of rate of substrate expenditure for cell maintenance and the “maximum” yield (i.e., hypothetical yield without cell maintenance processes) were estimated. Analysis of the maximum yield values based on mass–energy balance theory suggested that oxidation of the carboxylic acid side chains of EDTA by a monooxygenase had zero or low energetic efficiency. An Arrhenius equation with different values of Arrhenius parameters within different temperature ranges gave a good fit with the temperature dependence of both growth rate and biomass yield. Specific growth rates of both strains showed a more pronounced temperature dependence than did the cell yields. A possible kinetic mechanism was suggested which might be responsible for the modes of the temperature dependences of specific growth rate and yield that were found. The mechanism is based on a hypothetical key substance governing the metabolic flows, which is formed in a zero-order reaction and destroyed in a first-order reaction, both rate constants depending on temperature according to the Arrhenius law. 相似文献
997.
Growth of 2659 Atlantic cod Gadus morhua aged 4 to 9 years examined in Placentia Bay, Newfoundland, peaked in most cases in June and was at a minimum in October or November. Water temperature, partial fullness index ( I P ) and gonado‐somatic index ( I G ) explained between 31 and 52% of the monthly variability in growth. Temperature and I P of capelin Mallotus villosus had significant effects on growth of all age groups and explained most of the variance for ages 6–8 and 4–5 years, respectively. The I P of large invertebrates (ages 4 to 7 years), sandlance ( Ammodytes sp. age 6 years) and demersal fishes (age 9 years) had age‐specific effects in the model. Overall, amphipods, decapods and echinoderms dominated the Atlantic cod diet in most seasons, but fish consumption by Atlantic cod was high in June and July, particularly on capelin. The rapid increase in somatic mass during June and July occurred despite cold water temperatures ( < 3° C at 50 m) and moderate to high gonado‐somatic index. The findings of this study suggest that when food was not a limiting factor, growth tended to increase even when Atlantic cod occupied colder waters, but when food was limiting, the opposite may have occured. 相似文献
998.
Feather samples from 117 birds, representing 11 species, were examined for keratinophilic fungi. Of these, 63 (53.8%) were found to be positive. Nine species of fungi classified in 6 genera were isolated: Aphanoascus terreus (1.7%), Chrysosporium indicum (26.4%), Chrys. tropicum (11.1%) the Chrysosporium state of Aphanoascus spp. (2.5%), the Chrysosporium state of Arthroderma tuberculatum (3.4%), Ctenomyces serratus (0.8%), Gymnoascoideus petalosporus (7.6%), Malbranchea spp. (6.8%), Rollandina hyalinospora (6.8%). Passer domesticus (the house sparrow) harboured the largest varieties of keratinophilic fungi: the Chrysosporium state of Arthroderma tuberculatum was found to be associated with it selectively. The associations of these keratinophilic fungi with birds were non-clinical in nature. 相似文献
999.
1000.
Axial ligands of chloroplast cytochrome b-559: identification and requirement for a heme-cross-linked polypeptide structure 总被引:5,自引:0,他引:5
Optical, resonance Raman, and electron paramagnetic resonance spectroscopies have been used to characterize the ligands and spin state of the chloroplast cytochrome b-559. The protein was isolated from both maize and spinach in a low-potential form. The spectroscopic data indicate that the heme iron in both ferric and ferrous cytochrome b-559 is in its low-spin state and ligated in its fifth and sixth coordination positions by histidine nitrogens. Electron paramagnetic resonance data for the purified spinach cytochrome are in good agreement with those determined by Bergstr?m and V?nng?rd [Bergstr?m, J., & V?nng?rd, T. (1982) Biochim. Biophys. Acta 682, 452-456] for a low-potential membrane-bound form of cytochrome b-559. The g values of high-potential cytochrome b-559 are shifted from those of its low-potential forms; this shift is interpreted as arising from a deviation of the planes of the two axial histidine imidazole rings from a parallel orientation. The model is consistent with the physical data and may also account for the facility with which cytochrome b-559 can be converted between low- and high-potential forms. Recent biochemical and molecular biological data [Widger, W. R., Cramer, W. A., Hermodson, M., Meyer, D., & Gullifor, M. (1984) J. Biol. Chem. 259, 3870-3876; Herrmann, R. G., Alt, J., Schiller, D., Cramer, W. A., & Widger, W. R. (1984) FEBS Lett. 179, 239-244] have shown that two polypeptides, one with 83 residues and a second with 39 residues, most likely constitute the protein of the cytochrome.(ABSTRACT TRUNCATED AT 250 WORDS) 相似文献