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Targeting of porin to the outer membrane of Escherichia coli. Rate of trimer assembly and identification of a dimer intermediate 总被引:14,自引:0,他引:14
J Reid H Fung K Gehring P E Klebba H Nikaido 《The Journal of biological chemistry》1988,263(16):7753-7759
Porin, a transmembrane protein in the outer membrane of Escherichia coli, exists in a trimeric structure which is not dissociated during sodium dodecyl sulfate-polyacrylamide gel electrophoresis at 25 degrees C. This unusual stability was utilized in the study of the conformational changes which accompany the targeting of porin to the outer membrane. A delay of 16-44 s between completion of synthesis of a monomer and its assembly into a trimer was found from the ratio of monomers to trimers found in exponentially growing cells. Pulse-chase experiments showed that rapid processing of precursor OmpF molecules was followed by assembly into sodium dodecyl sulfate-resistant oligomers with a half-time of 20 s at 30 degrees C. An intermediate in assembly was isolated by immunoprecipitation and sodium dodecyl sulfate-polyacrylamide gel electrophoresis below 10 degrees C and was identified as a metastable dimer. 相似文献
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Monoclonal anti-mouse macrophage antibodies recognize the globular portions of C1q, a subcomponent of the first component of complement 总被引:1,自引:0,他引:1
H P Heinz H Dlugonska E Rüde M Loos 《Journal of immunology (Baltimore, Md. : 1950)》1984,133(1):400-404
One of seven monoclonal antibodies generated against mouse macrophages (M phi) was found to recognize isolated heterologous C1q. This antibody was shown to be cytotoxic and to react in a strain-independent way with mouse M phi derived from bone marrow cells as well as with M phi from the peritoneal cavity; it did not react, however, with mouse granulocytes, thymocytes, or T and B lymphocytes. The hemolytic activity of fluid phase C1q was inhibited to 50% at a 2 X 10(-4) dilution of hybridoma supernatant, whereas a 100-fold higher concentration was required to inhibit C1q bound to immune complexes ( EAC1q ) to the same extent. It was demonstrated that this antibody recognizes the isolated globular, Fc-binding portions of the C1q molecule and reacts with the A and B chains. Because M phi have been shown to synthesize C1q, the Fc-recognizing subcomponent of the first component of complement, evidence was provided that endogeneous C1q can serve as an Fc receptor on M phi during secretion. This fact was demonstrated by a dose-dependent inhibition of Fc-receptor activity for EIgG by the F(ab')2 fragment of this monoclonal antibody. These experiments further support the concept that C1q produced by M phi functions on the surface as an Fc-recognizing molecule before it is released and incorporated into the macromolecular complex of serum C1. 相似文献
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BACKGROUND: Clinical outcome predictions in phase III studies are mostly derived for patient groups, but not for individual patients, although individualised predictions are an ultimate goal to permit a personalised fine tuning of therapy. This may permit earlier application of target therapies, minimise general damage to the organism, and result in improved complete remission rates in malignant diseases. METHODS: In this study, Lymphochip cDNA microarray gene expression results of DLBCL patients, from a published prospective meta-analysis study on the prediction of group prognosis, were analysed for individualised predictions using a nonstatistical data pattern classification approach. The training set was comprised of the same 160 DLBCL patients as in the prognosis study, with the validation set of 80 patients remaining unknown to the learning process. This permits the assessment of prospective classifier performance towards unknown patients. RESULTS: Pretherapeutic predictions for the training and validation set patients were correct in 98.1% and 78.3% of the cases for nonsurvival and in 67.3% and 45.3% for survival. The discriminatory data pattern consisted of 14 known and 10 unknown gene products. CONCLUSIONS: The better than 95% correct pretherapeutic prediction for about one-half of the ultimately nonsurviving high-risk patients of the training set is promising for clinical considerations about individualised therapy in such cases. Reliable individualised survival predictions are not possible with the information content of the present dataset. It seems necessary to investigate additional gene products, since survival may significantly depend on non-lymphocyte-associated genes that escape to the lymphocyte-oriented Lymphochip gene activation analysis. 相似文献
8.
The accumulation of mature RNA for the Xenopus laevis ribosomal protein L1 is controlled at the level of splicing and turnover of the precursor RNA. 总被引:17,自引:5,他引:12 下载免费PDF全文
A specific control regulates, at the level of RNA splicing, the expression of the L1 ribosomal protein gene in Xenopus laevis. Under particular conditions, which can be summarized as an excess of free L1 protein, a precursor RNA which still contains two of the nine introns of the L1 gene accumulates. In addition to the splicing block the two intron regions undergo specific endonucleolytic cleavages which produce abortive truncated molecules. The accumulation of mature L1 RNA therefore results from the regulation of the nuclear stability of its precursor RNA. We propose that a block to splicing can permit the attack of specific intron regions by nucleases which destabilize the pre-mRNA in the nucleus. Therefore the efficiency of splicing could indirectly control the stability of the pre-mRNA. 相似文献
9.
Heinz W. Kunz Andrea L. Cortese Hassett Tetsuo Inomata D. N. Misra Thomas J. Gill III 《Immunogenetics》1989,30(3):181-187
A new antigenic system in the rat homologous to theQa/TL antigen system in the mouse has been characterized. It was detected by antibodies raised in donor-recipient combinations
that were matched for theRT1. A, B, D, E loci in the major histocompatibility complex (MHC): (R11×BN)F1 anti-BN.1L(LEW), (R18×BN)F1 anti-BN.1L, and BN.1LV1(F344) anti-BN.1L. Absorption analyses using these antisera and a variety of inbred, congenic and
recombinant strains identified three alleles,RT1.G
a
,G
b
,G
c
, of whichG
c
is a null allele. The strain distribution of these alleles was determined, using 37 strains of rats representative of all
of the prototypic haplotypes and a number of congenic and recombinant strains. The use of the congenic and recombinant strains
showed that theRT1.G locus was linked to the MHC and that the most probable gene order wasA-E-G. Testcross analysis showed that the map distance betweenA andG was 1.4 cM(4/285 recombinants). The RT1.G antigen has a heavy chain ofM
r 46 000 and is present on both T and B cells. 相似文献
10.
Ken S. Dhalla Pallab K. Ganguly Heinz Rupp Robert E. Beamish Naranjan S. Dhalla 《Molecular and cellular biochemistry》1989,87(1):85-92
Using the reverse phase high-performance liquid chromatography (HPLC) with mobile phases composed of simple acids, we have developed an assay technique for the measurement of adrenolutin, one of the oxidation products of catecholamines, in rat plasma. Ion-pairing chromatography permits the separation and quantitation of plasma adrenolutin (M) in a linear manner. Sample preparation involved the precipitation of plasma proteins with perchloric acid and it is easier to handle a large number of samples at a time. However, we were unable to demonstrate the presence of adrenochrome, another oxidation product of catecholamines, in plasma since adrenochrome was rapidly destroyed in acid as well as in blood and was quickly changed, into adrenolutin. Adrenolutin peak in HPLC was confirmed by 1) the retention time; 2) co-injection of adrenolutin and; 3) the appearance of 3H-adrenolutin after injection of 3H-norepinephrine. Administration of different catecholamines as well as adrenochrome and adrenolutin in rats also increased the level of adrenolutin in plasma. Adrenolutin was found to be present in plasma in other species including dog, rabbit and pig. High level of adrenolutin, which may represent total concentration of aminolutin in plasma, suggests the presence of an efficient mechanism for the oxidation of catecholamines under in vivo conditions. 相似文献