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1.
A monoclonal antibody to bovine brain inositol monophosphatase. Immunoaffinity purification of the brain and kidney enzymes and evidence for their structural identity. 总被引:1,自引:1,他引:0 下载免费PDF全文
A monoclonal IgG2b(K) antibody, G-2A4, has been generated against bovine brain myo-inositol monophosphatase (EC 3.1.3.25). The identity of the antigen recognized by the antibody was established by using e.l.i.s.a. and Western blotting procedures, and by immunoprecipitation of enzyme activity from crude brain supernatant. In addition, the hydrolysis of Ins1P by crude brain extract was inhibited by up to 83% by the pure antibody. Under identical conditions, the hydrolysis of Ins(1,4)P2 was unaffected. An immunoadsorbent column containing monoclonal antibody G-2A4 covalently attached to CNBr-activated Sepharose 4B has been used for rapid purification of the brain enzyme. Elution conditions have been optimized to allow isolation of the enzyme in high yield (54%) with full retention of column-binding capacity. The enzyme was electrophoretically homogeneous, Mr 30,000 and of higher specific activity than that purified conventionally. Chromatography of the pure enzyme on high resolution ion-exchange columns revealed some charge heterogeneity, possibly indicative of some type of post-translational modification. The immunoadsorbent column has also been used to purify the bovine kidney cortex enzyme to homogeneity. Partial proteolytic fragmentation patterns of the brain and kidney enzymes using endoprotease glu-C were identical, suggesting that they are almost certainly products of the same gene. 相似文献
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The dephosphorylation of inositol 1,4-bisphosphate to inositol in liver and brain involves two distinct Li+-sensitive enzymes and proceeds via inositol 4-phosphate. 总被引:13,自引:7,他引:6 下载免费PDF全文
C I Ragan K J Watling N S Gee S Aspley R G Jackson G G Reid R Baker D C Billington R J Barnaby P D Leeson 《The Biochemical journal》1988,249(1):143-148
1. Hydrolysis of both enantiomers of inositol 1-phosphate and both enantiomers of inositol 4-phosphate to inositol is inhibited by LiCl in liver and brain. 2. The phosphatase activity is predominantly soluble. 3. Inositol 1,4-bisphosphate is also hydrolysed by the soluble fraction of liver and brain. 4. Bisphosphatase activity is inhibited by LiCl, but is less sensitive than monophosphatase activity. 5. The product of bisphosphatase in liver and brain is inositol 4-phosphate. 相似文献
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R E Weller R L Buschbom H A Ragan J F Baer C A Malaga 《Journal of medical primatology》1990,19(5):439-446
A positive linear relationship was found between total calcium and albumin and between total calcium and total protein in the serum of 205 owl monkeys. Adjustment formulas for calcium were derived: adjusted calcium (mg/dl) = measured calcium (mg/dl) - 0.84 [albumin (g/dl)] + 3.8 and adjusted calcium (mg/dl) = measured calcium (mg/dl) - 0.82 [total protein (g/dl)] + 6.5. Adjusted calcium calculations for monkeys based on albumin concentration were similar to those for man and dogs, but different from those based on total protein. 相似文献
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Kinley Ragan Jan Schipper Heather L. Bateman Sharon J. Hall 《The Journal of wildlife management》2023,87(1):e22322
Wildlife populations in semi-arid regions are increasingly challenged by human activities and dependent on the connectivity of riparian corridors for access to surface water. The Madrean Archipelago is a biodiversity hotspot along the arid United States–Mexico borderlands that support both Neotropical and Nearctic wildlife. Infrastructure development (e.g., the border wall and the expansion of Mexican Federal Highway 2) in this region inhibits wildlife movement along the transnational mountain archipelago by disconnecting habitat. To explore the relationship between habitat variables and mammal use of riparian corridors in northern Sonora, Mexico, we collected data from 19 motion-sensitive cameras between October 2018 and April 2019 and used single-season occupancy models and Royle-Nichols abundance estimation models to analyze our data. We recorded 21 species of mammals, including the first sighting of jaguar (Panthera onca) in this region in 25 years. River characteristics (distance from river, riparian corridor width, water availability), remoteness (distance from highway, productivity, elevation), and topographic variety (vertical elevation difference) influenced patterns of occupancy probability and estimated abundance of mammals >1 kg, but the strength and direction of these relationships varied by species. Additionally, intermittently wet desert washes were comparable in species richness to the perennial system. These results highlight the importance of examining physical and biological aspects of habitat. This is especially true when identifying corridors where mitigation structures should be placed to improve wildlife connectivity in biodiversity hotspots like the Madrean Archipelago and semi-arid ecosystems worldwide. 相似文献
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Methods for estimating the absolute polyphenol content of the brown algae Ascophyllum nodosum (L.) Le Jol. and Fucus vesiculosus (L.) have been developed and tested. Polyphenols were extracted almost quantitatively from Ascophyllum nodosum using aqueous acetone, whereas this procedure was somewhat less efficient with Fucus vesiculosus. Colorimetric methods based on the Folin-Denis reagent, Brentamine Fast Red 2G Salt, and vanillin-H2SO4 were applied to acetone-free extracts for determination of polyphenol content relative to suitable reference compounds. Gravimetric methods based on hide powder and on haemoglobin were employed to derive ‘estimation factors’ (EFs) which allow calculation of the absolute polyphenol content from the relative polyphenol content. The values calculated for absolute polyphenol content are considered to be reasonably accurate, despite imprecisions in the methods and despite often large standard deviations, and re-emphasize the potential physiological and ecological significance of brown algal polyphenols. Although the precise EFs calculated here are not valid for other brown algae, the methods are considered to be generally applicable to other Phaeophyceae. 相似文献
9.
Purification and properties of L-3-glycerophosphate dehydrogenase from pig brain mitochondria. 总被引:2,自引:2,他引:0 下载免费PDF全文
L-3-Glycerophosphate dehydrogenase (EC 1.1.99.5) was purified from pig brain mitochondria by extraction with deoxycholate, ion-exchange chromatography and (NH4)2SO4 fractionation in cholate, and preparative isoelectric focusing in Triton X-100. Sodium dodecyl sulphate/polyacrylamide gel electrophoresis shows that the purified enzyme consists of a single subunit of mol.wt. 75 000. The enzyme contains non-covalently bound FAD and low concentrations of iron and acid labile sulphide. No substrate reducible e.p.r. signals were detected. The conditions of purification, particularly the isoelectric focusing step, lead to considerable loss of FAD and possibly iron-sulphur centres. It is therefore not possible to decide with certainty whether the enzyme is a flavoprotein or a ferroflavoprotein. The enzyme catalyses the oxidation of L-3-glycerophosphate by a variety of electron acceptors, including ubiquinone analogues. A number if compounds known to inhibit ubiquinone oxidoreduction by other enzymes of the respiratory chain failed to inhibit L-3-glycerophosphate dehydrogenase, except at very high concentrations. 相似文献
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