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Human platelets contain an enzyme that catalyzes CoA-independent release of arachidonic acid from phosphatidylcholine with concomitant incorporation into plasmenylethanolamine. Addition of lysoplasmenylethanolamine (10-80 microM) to a crude membrane preparation of prelabeled platelets (0.24 mg of protein/ml) induces transfer of [3H]arachidonate from endogenous phosphatidylcholine to lysoplasmenylethanolamine (0.8 nmol of arachidonic acid/min/mg of protein). The transacylation reaction occurs in the absence of Ca2+, has a broad pH optimum from 7 to 8, is not affected by excess unlabeled arachidonic acid, and is inhibited by N-ethylmaleimide (0.2 mM) and Triton X-100 (0.1 mg/ml). The enzyme shows a high specificity toward the acyl donor (phosphatidylcholine), transfers fatty acids in the order: arachidonic greater than eicosatrienoic greater than oleic, and preferentially acylates lysoplasmenylethanolamine but also other lysophosphatides (lysophosphatidylethanolamine greater than lysophosphatidylserine greater than lysophosphatidylinositol = 0). Platelet acyltransferase, on the other hand, acylates ethanolamine lysophosphatides with free arachidonic acid in the order: lysophosphatidyl-ethanolamine greater than lysoplasmenylethanolamine. These results suggest that a distinct acylation mechanism exists for introduction of arachidonic acid into plasmalogen phosphatides. In stimulated platelets, the transacylase may play an additional role in the controlled release of esterified arachidonic acid for synthesis of the biologically active oxygenated metabolites. 相似文献
43.
Photolabile derivatives of bile salts. Synthesis and suitability for photoaffinity labeling 总被引:4,自引:0,他引:4
In an approach to the identification of bile salt-binding carriers, the photoactivable bile acid derivatives A) 3 beta-azido, 7 alpha,12 alpha-dihydroxy-5 beta-cholan-24-oic acid, B) 7,7-azo-3 alpha,12 alpha-dihydroxy-5 beta-cholan-24-oic acid, and C) 11 xi-azido-12-oxo-3 alpha,7 alpha-dihydroxy-5 beta-cholan-24-oic acid were synthesized in unconjugated and taurine-conjugated form. Photolysis of the 3 beta-azido derivatives was studied using a light source with a maximum emission at 300 nm and established a half-life time of 18.5 min. The photochemistry of the 7,7-azo derivatives was investigated using light with a maximum at 350 nm and had a half-life time of 2.2 min. The 11 xi-azido-12-oxo derivatives were photolyzed with light having a maximum at 300 nm resulting in a half-life time of 8.5 min. The suitability of the 7,7-azo derivatives for photoaffinity labeling was demonstrated by photolyses in 14C-labeled methanol and acetonitrile. The generated carbene reacted with the solvents under covalent bond formation of 6 to 12%. The efficiency of all synthesized photolabile derivatives for photoaffinity labeling of bile salt binding proteins was demonstrated. 相似文献
44.
Comparative analysis of the 5''-end regions of two repressible acid phosphatase genes in Saccharomyces cerevisiae. 总被引:21,自引:12,他引:9 下载免费PDF全文
The nucleotide sequence of 5'-noncoding and N-terminal coding regions of two coordinately regulated, repressible acid phosphatase genes from Saccharomyces cerevisiae were determined. These unlinked genes encode different, but structurally related polypeptides of molecular weights 60,000 and 56,000. The DNA sequences of their 5'-flanking regions show stretches of extensive homology upstream of, and surrounding, a "TATA" sequence and in a region in which heterogeneous 5' ends of the p60 mRNA were mapped. The predicted amino acid sequences encoded by the N-terminal regions of both genes were confirmed by determination of the amino acid sequence of the native exocellular acid phosphatase and the partial sequence of the presecretory polypeptide synthesized in a cell-free protein synthesizing system. The N-terminal region of the p60 polypeptide was shown to be characterized by a hydrophobic 17-amino acid signal polypeptide which is absent in the native exocellular protein and thought to be necessary for acid phosphatase secretion. 相似文献
45.
Corn (Zea mays L.) and sorghum (Sorghum vulgare, Pers.) plants were grown in a vermiculite-gravel mixture in controlled environment chambers until they were 40 days old. Water was withheld until they were severely wilted, and they were then rewatered. During drying and after rewatering stomatal resistance was measured with a diffusion porometer each morning, and water saturation deficit and water potential were measured on leaf samples. The average resistance of the lower epidermis of well watered plants was lower for corn than for sorghum. When water stress developed, the stomata began to close at a higher water potential in corn than in sorghum. The stomata of both species began to reopen normally soon after the wilted plants were rewatered, and on the 2nd day the leaf resistances were nearly as low as those of the controls. The average leaf water potential of well watered corn was −4.5 bars; that of sorghum, −6.4 bars. The lowest leaf water potential in stressed corn was −12.8 bars at a water saturation deficit of 45%. The lowest leaf water potential in stressed sorghum was −15.7 bars, but the water saturation deficit was only 29%. At these values the leaves of both species were tightly rolled or folded and some injury was apparent. Thus, although the average leaf resistance of corn is little lower than that of sorghum, corn loses much more of its water before the stomata are fully closed than does sorghum. The smaller reduction in water content of sorghum for a given reduction in leaf water potential is characteristic of drought-resistant species. 相似文献
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Lactate dehydrogenase subunits B and A are produced by genes at separate loci. LDH-1, the most anodal of the five isozymes observed after gel electrophoresis, is composed of four B subunits. It has recently been shown that the LDH-1s of most primates are electrophoretically the same. N. coucang (slow loris) is one of the exceptions, possessing an LDH-1 which migrates more slowly than that common to most other primates. We have observed in some members of N. coucang a band at the site of the common primate LDH-1 in addition to the LDH-1 normally present. Since one of the animals in which this observation was made was heterozygous at the LDH B locus, we concluded that in N. coucang two gene loci coding for the B polypeptide are probably present.This investigation was supported in part by contract AF 29 (600)-5587 and NSF grant GB-7426. 相似文献
50.
Nature of the Conversion of Ficus carica Variety Kadota Ficin Component D to Component C. Some Physicochemical Properties of Components C and D 总被引:3,自引:3,他引:0 下载免费PDF全文
Component C can be formed from component D under the experimental conditions used during purification of Ficus carica variety Kadota latex. By use of the inhibitor, sodium p-chloromercuribenzoate, the 2 components have been purified to chromatographic homogeneity. The 2 components have identical molecular weights and amino acid composition. The only difference found between the 2 components is the presence of 3 to 6 more amide groups in component D than in component C. There also appears to be a conformational difference between the 2 since component C is not as acidic, with respect to component D, as would be expected from the comparative amide contents. Conformational differences between the 2 are also indicated by the chromatographic behavior of the 2 in the presence and absence of sodium-p-chloromercuribenzoate. 相似文献