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1.
Inactivation of C3a by a monocarboxypeptidase present in culture supernatants of stimulated guinea pig peritoneal macrophages 总被引:1,自引:0,他引:1
G Kreuzpaintner B Damerau B Zimmermann T H Plummer V Brade 《Journal of immunology (Baltimore, Md. : 1950)》1986,136(9):3384-3389
Hog C3a, as well as its derivative C3a-desArg were not found to act cytotoxically on starch gel-induced guinea pig peritoneal macrophages. Likewise, neither peptide significantly modified the secretion of N-acetyl-beta-D-glucosaminidase from these cells. However, C3a rapidly lost its spasmogenic activity during incubation in serum-free macrophage cultures and less rapidly in cellfree supernatants collected from cultured macrophages. The following results indicate that C3a is converted into its spasmogenically inactive derivative C3a-desArg by a macrophage-derived monocarboxypeptidase. The inactivated C3a product does not differ from native C3a in sodium dodecyl sulfate-polyacrylamide gel electrophoresis; it elutes from CM cellulose in the same position as purified C3a-desArg; and it is devoid of the carboxyl-terminal arginyl residue of C3a, but still contains the carboxyl-terminal sequence of C3a-desArg as determined by analysis after treatment with carboxypeptidases B or Y. Furthermore, inactivation of C3a in supernatants of macrophage cultures is completely blocked by the specific carboxypeptidase inhibitors guanidinopropylsuccinic acid and 2-mercaptomethyl-3-guanidinoethylthiopropanoic acid in final concentrations of 10 mM and 2.1 mM, respectively. The monocarboxypeptidase is apparently supplied by biosynthesis of new material but is not stored as a preformed enzyme because cycloheximide markedly inhibits its expression. 相似文献
2.
3.
Four equine herpesviruses (equine abortion virus, equine herpesvirus types 2 and 3, and equine cytomegalovirus) were compared. The equine abortion virus did not cross-neutralize with any of the other viruses, but the other three did show varying degrees of cross-neutralization among themselves. Equine abortion virus grew more quickly in tissue cultures than did the others, and attained higher titers of infectivity in the culture fluid; it also formed plaques in a wider range of tissue culture species, although the other three were not specific for one tissue culture system only, in that they would multiply in rabbit and cat kidney cultures. The densities of the deoxyribonucleic acids of all four viruses were in the range 1.716 to 1.717 g/ml (a guanine plus cytosine content of 57 to 58%). Taxonomic separation, as a distinct serotype, of equine abortion virus from the other herpesviruses seems to be justified. The other three are closely related to one another. They should perhaps be regarded as separate viruses and termed horse herpesviruses types 2, 3, and 4, although an alternative view would be to regard them as variants of a single virus type. The question of whether types 2, 3, and 4, or any other herpesviruses, should be placed in a phylogenetically distinct subgroup, known as cytomegaloviruses, is a moot point. 相似文献
4.
T H Plummer 《The Journal of biological chemistry》1968,243(22):5961-5966
5.
Genetic variation at the Major Histocompatibility Complex locus DQ beta was
analyzed in 233 beluga whales (Delphinapterus leucas) from seven
populations: St. Lawrence Estuary, eastern Beaufort Sea, eastern Chukchi
Sea, western Hudson Bay, eastern Hudson Bay, southeastern Baffin Island,
and High Arctic and in 12 narwhals (Monodon monoceros) sympatric with the
High Arctic beluga population. Variation was assessed by amplification of
the exon coding for the peptide binding region via the polymerase chain
reaction, followed by either cloning and DNA sequencing or single-stranded
conformation polymorphism analysis. Five alleles were found across the
beluga populations and one in the narwhal. Pairwise comparisons of these
alleles showed a 5:1 ratio of nonsynonymous to synonymous substitutions per
site leading to eight amino acid differences, five of which were
nonconservative substitutions, centered around positions previously shown
to be important for peptide binding. Although the amount of allelic
variation is low when compared with terrestrial mammals, the nature of the
substitutions in the peptide binding sites indicates an important role for
the DQ beta locus in the cellular immune response of beluga whales.
Comparisons of allele frequencies among populations show the High Arctic
population to be different (P < or = .005) from the other beluga
populations surveyed. In these other populations an allele, Dele-DQ
beta*0101-2, was found in 98% of the animals, while in the High Arctic it
was found in only 52% of the animals. Two other alleles were found at high
frequencies in the High Arctic population, one being very similar to the
single allele found in narwhal.
相似文献
6.
Margaret S.Y. Chai Charles A. Reavill Cecelio J. Vidal David T. Plummer 《Neurochemistry international》1981,3(5):311-321
Extraction of the white matter of pig brain with EDTA, lysolecithin or Triton X-100 gave poor yields of soluble acetylcholinesterase although these agents had proved effective at solubilizing the enzyme in the grey matter. This finding, together with the observation that the strong detergent sodium deoxycholate, was needed to solubilize the enzyme, shows that it is more difficult to remove acetylcholinesterase from the white matter of brain than from the grey. This could mean that the enzyme in the white matter is more firmly bound to the membrane than the enzyme in the grey matter.The difference in binding of the enzyme from the two regions of the brain is also reflected in the affinity chromatography experiments which showed a lower recovery for the acetylcholinesterase of white matter compared with the enzyme from grey matter.Starch-block electrophoresis of acetylcholinesterase showed a single negatively charged peak of activity for both the naturally soluble and the deoxycholate solubilized preparations. The presence of only one form on electrophoresis suggests that the molecular species of acetylcholinesterase do not arise from differences in charge.Sucrose density gradient centrifugation of the two preparations from white matter gave a single peak of activity with a sedimentation constant of about 10 S. This corresponds closely to the major species of molecular weight 260,000 detected by gradient gel electrophoresis. Other forms detected in both enzyme preparations by gradient gel electrophoresis were species with molecular weights of 660,000, 180,000, 130,000 and 115,000. The significance of these species in terms of the formation of oligomers is discussed.A comparison was made with the corresponding preparations of acetylcholinesterase from the grey matter and the results showed that acetylcholinesterase from the white and grey matter of pig brain were very similar. The exception to this was the species with a molecular weight of 68,000 which was present in the grey but not the white matter of pig brain. 相似文献
7.
Immunoreactive beta-melanocyte-stimulating hormone (beta-MSH) was measured in the plasma of 19 patients with hypopituitarism and in the cerebrospinal fluid (CSF) of five of these patients. In neither plasma nor CSF were the beta-MSH concentrations significantly different from those in normal controls. These observations raise the possibility that beta-MSH may be produced by and secreted from neural tissue; this is supported by the findings of beta-MSH in high concentrations in many parts of the brain. 相似文献
8.
F M Melewicz J M Plummer H L Spiegelberg 《Journal of immunology (Baltimore, Md. : 1950)》1982,129(2):563-569
Fc receptors for IgE (Fc epsilon R) on human peripheral blood lymphocytes and monocytes and cultured lymphoblastoid and macrophage-like cell lines were compared with respect to: 1) binding affinity for radiolabeled IgE, 2) inhibition of IgE-specific rosette formation and inhibition of binding of radiolabeled IgE by an antiserum raised against Fc epsilon R isolated from a lymphoblastoid cell line, and 3) m.w. of radiolabeled cell surface proteins precipitated with the anti-Fc epsilon R serum. Scatchard analysis of 125I-IgE binding to lymphocytes, monocytes, and their corresponding cell lines showed biphasic binding curves with all cell types, from which 2 binding affinities were calculated to be KA = 6.2 +/- 1.1 and 2.0 +/- 0.5 x 10(7) M-1. The anti-Fc epsilon R serum inhibited both IgE rosette formation and binding of radiolabeled IgE by lymphocytes and monocytes but did not inhibit IgE rosettes formed by basophils. The inhibitory activity of the anti-Fc epsilon R serum could be absorbed with Fc epsilon R(+) but not with Fc epsilon R(-) cell lines. The anti-Fc epsilon R serum precipitated 2 peptides having m.w. of approximately 47,000 and 23,000 daltons from lysates of both cell surface-labeled lymphocyte and macrophage cell lines. These data indicate that Fc epsilon R on normal lymphocytes and monocytes, as well as on cultured lymphoblastoid and macrophage-like cells, are related structurally, since they share antigenic determinants, bind IgE with a similar affinity, and have similar m.w. However, they differ in all 3 parameters from Fc epsilon R on basophilic granulocytes. 相似文献
9.
CNDO/2 studies on the conformation of the chain of lecithin indicated a strong preference for a gauche-gauche arrangement about the phosphodiester group. Folding the chain about and 4 was energetically very favorable. Hydration of the same segment revealed three levels of water-binding energies. The ion-dipole interactions of water and the choline moiety were energetically non-substantial. In contrast, binding of water to the unesterified phosphate oxygens produced the highest enthalpies. Attachment of water to the esterified phosphate oxygens or the ester oxygens of the chain resulted in intermediate binding strengths. By investigating complete incorporation of nine water molecules into a chosen lipid structure, a plausible lecithin-water geometry was deduced for a liquid crystalline system. 相似文献
10.
A continuous spectrophotometric assay for human plasma carboxypeptidase N utilizing furylacryloyl-alanyl-lysine is described. Synthesis was made by use of 9-(2-sulfo)fluorenylmethyloxycarbonyl (Sulfmoc) chloride as the N-?-amino-blocking group for lysine. The substrate has the advantage of containing a chromophore which allows difference measurements above 324 nm. The kinetic parameters Km and have been determined for furylacryloyl-alanyl-lysine and -arginine. Difference measurements were related to micromoles of lysine or arginine released and were expressed as units. 相似文献