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1.
We have isolated cDNAs for carcinoembryonic antigen (CEA) and for a normal cross-reacting antigen (NCA) and report here their nucleotide and derived amino acid sequences. Our data show that both the CEA and NCA polypeptides are organized into extracellular domains, some with cysteine-linked loops, that share extensive sequence homology (approximately 78% overall) with each other and appear similar to immunoglobulin superfamily members. A major difference between the two apoproteins is the presence of a single loop-domain in NCA compared to three tandemly repeated loop-domains in CEA. Sequence comparisons between the extracellular domains of CEA and NCA show that the N-terminal and adjacent loop domains of each apoprotein have high homology (85-90%) to each other, while comparison of loop-domain regions reveals a possible nonrandom distribution of base changes and altered amino acids near certain cysteine residues that are inferred to be involved in forming disulfide loops. Both apoproteins show high identity in their hydrophobic C-termini that are reminiscent of the type of transmembrane tails seen in proteins that potentiate signal transduction. These findings, coupled with distinct expression profiles of CEA and NCA mRNAs, suggest that these apoproteins may function as unique cell-surface molecules mediating cell-specific interactions in normal and neoplastic cells.  相似文献   

2.
Nonspecific cross-reacting antigen (NCA) is a highly glycosylated membrane protein which is immunologically and structurally related to carcinoembryonic antigen, an important tumor-associated antigen. Two glycoforms of NCA were purified from a single liver metastasis of a colonic carcinoma and characterized with respect to their primary sequence and position of glycosylation sites. The two glycoforms (designated TEX (tumor-extracted antigen), Mr 75,000, and NCA, Mr 45,000) each showed a deglycosylated Mr of 35,000 and yielded identical peptide maps. The structural characterization of TEX and NCA and the assignment of glycosylation sites was performed by fast atom bombardment mass spectrometry and microsequence analysis of the resulting peptides. This approach showed that TEX and NCA were identical with respect to primary sequence and provided direct evidence that 11 of the 12 predicted asparagine-linked glycosylation sites were glycosylated in both TEX and NCA. Indirect evidence was obtained for glycosylation at the other site. Both glycoforms also contain ethanolamine linked to Gly-286, a finding consistent with the conclusion that these proteins are anchored to the plasma membrane through a glycosyl-phosphatidylinositol tail. The large difference in the molecular weights of glycosylated TEX and NCA suggests significant variations in their oligosaccharide structures.  相似文献   

3.
Despite its being weaker base poly(2-vinylpyridine) polymerized DL -β-phenylalanine NCA at a much faster rate than pyridine and α-picoline. Poly(2-vinylpyridine) adsorbs NCA by hydrogen bonding with the cooperation of a few pyridine groups. This results in a high local concentration of NCA. The syndiotactic configuration of pyridine group seemed to be least suitable for the cooperative hydrogen bonding. Adsorbed NCA is activated to form an “activated” NCA which in turn reacts with an NCA adsorbed on the same polymer chain. Since the polymer chain is flexible, this intramolecular reaction takes place frequently, resulting in the acceleration of polymerization. The intramolecular reaction along the polymer chain is dependent on the degree of polymerization of polymer catalyst. A suitable model was proposed for the intramolecular reaction to explain the effect of degree of polymerization.  相似文献   

4.
The surfaces of non-geniculate coralline algae (NCA) are known to induce the settlement and metamorphosis of disparate marine taxa. In this study we investigate the responsiveness of larvae of Herdmania curvata (Ascidiacea: Stolidobranchia) to three species of NCA (Neogoniolithon brassica-florida, Hydrolithon onkodes, and Lithothamnium prolifer) that cohabit the slope and crest of Heron Reef, Great Barrier Reef. H. curvata larvae were first exposed to these NCA at or within 2 h of hatching, which is 1 to 2 h prior to attaining competence, and then cultured continuously with the NCA for 12 to 14 h. Rates of settlement and metamorphosis of H. curvata cultured in laboratory chambers in the presence of the different NCA were significantly lower than spontaneous rates in seawater. The limited settlement in treatments containing NCA were confined entirely to the chamber periphery, and settlement never occurred on the surface of the NCA. The inhibitory effect was dose-dependent and was stronger in N. brassica-florida and H. onkodes than in L. prolifer. Larvae that did not settle in treatments with NCA had rounded anterior trunks and, in extreme cases, kinked tails with rounded and dissociated tail muscle cells. In some individuals, we observed the anterior chemosensory papillae being sloughed off the larval body. Morphological analysis of trunk ectodermal and mesenchymal nuclei of larvae cultured in the presence of the NCA revealed that general necrotic cell death was occurring. Importantly, H. curvata larvae that were exposed to NCA could not subsequently be induced to metamorphose in KCl-elevated seawater, whereas larvae not exposed to NCA metamorphosed at high rates in KCl-elevated seawater.  相似文献   

5.
The addition reaction to N-methyl-(S)-alanine or N-methyl-(S)-phenylalanine N-car-boxyanhydride (NCA) of 3-methyl-5-substituted hydantoin (HDT) catalyzed by a tertiary amine was investigated as a model reaction for the propagation reaction of NCA according to the activated-NCA mechanism. Several activated HDTs having the (S)-configuration of the asymmetric carbon atom were found to react more rapidly than their activated enantiomers. This experimental result indicates that the enantiomer selection by terminal-unit control takes place in the propagation reaction according to the activated-NCA mechanism in which an activated NCA is added to a terminal acylated NCA ring of the growing chain. The enantiomer excess of the HDT recovered from the reaction mixture of N-methyl-(S)-phenylalanine NCA and racemic HDTs activated by a tertiary amine was determined. The extent of the enantiomer selection in the polymerization was found to be 3–10 times as large as that in the model reaction. From these results, it was concluded that the chirality of the penultimate unit, as well as that of the terminal NCA ring, plays an important role in determining the enantiomer selection in the NCA polymerization.  相似文献   

6.
γ-Aminobutyric acid (GABA) receptors are postsynaptic membrane protein complexes that are important not only in the regulation of the nervous system but also as targets of drugs and insecticides. We synthesized a photoreactive straight-chain noncompetitive antagonist (NCA), 2-nitro-4-[3-(trifluoromethyl)-3H-diazirin-3-yl]phenyl 4-(4-methoxycarbonyl-1-butynyl)benzoate (NMB), to probe the NCA binding site. Our data show that this probe labels the NCA site and demonstrate that the NCA insecticide fipronil binds at a site distinct from that of other NCAs, such as picrotoxinin and 4′-ethynyl-4-n-propylbicycloorthobenzoate. The unique molecule NMB will be useful in identifying the cross-linking site of straight-chain NCAs in GABA receptors and mapping allosteric binding sites. Such studies should provide invaluable information in designing novel NCAs.  相似文献   

7.
As a step to investigate the cell adhesion mechanism and physiological roles of two CD66 antigens in human neutrophils, carcinoembryonic antigen gene family member 6 (CGM6, CD66b) and nonspecific cross-reacting antigen (NCA, CD66c), we prepared their soluble recombinant forms in silkworm larvae. Each cDNA fragment for CGM6 and NCA was ligated into the transfer vector pBK283 after modification to encode the protein lacking the membrane anchor. The resultant vectors were introduced to theBombyx morinuclear polyhedrosis virus, with which silkworm larvae were infected. Recombinant proteins secreted into the hemolymph of larvae at concentrations up to 1.3 mg/ml were purified by cation exchange followed by gel filtration or antibody affinity chromatography. The smaller apparent masses of the antigens compared with those of the native antigens appeared to be primarily due to incomplete glycosylation. Both recombinant antigens are quite similar to the corresponding native antigens in terms of the antigenic reactivity against a panel of CD66 monoclonal antibodies. In addition, the recombinant CGM6 and NCA exhibited cell binding activity against CHO cells expressing NCA and CGM6, respectively. Thus the two biologically active recombinant CD66 antigens prepared in large quantities in silkworm larvae should be useful for their functional studies, and our present system will be available for the production and purification of other carcinoembryonic antigen family members, whose biological functions are also unknown.  相似文献   

8.
Immobilized carcinoembryonic antigen (CEA) and non-specific crossreacting antigen (NCA) bound 3 strains of E. coli of human origin. The binding was dose dependent, saturable, and of high avidity. Binding of the bacteria to CEA and NCA was completely abolished in the presence of 10 mM alpha-methyl D-mannopyranoside. Bacteria did not bind to concanavalin A. In addition, binding to deglycosylated CEA was either absent or significantly reduced. These findings indicate that the E. coli strains bind to D-mannosyl residues in CEA and NCA. Considering the tissue distribution of CEA (brush border of colonic epithelium) and NCA (granulocytes), these glycoproteins may be involved in the recognition of bacteria.  相似文献   

9.
Peroxynitrite, formed by the reaction between nitric oxide and superoxide, has been shown to induce protein nitration, which compromises protein function. We hypothesized that peroxynitrite may regulate cytokine function during inflammation. To test this hypothesis, the neutrophil chemotactic activity (NCA) of interleukin-8 (IL-8) incubated with peroxynitrite was evaluated. Peroxynitrite attenuated IL-8 NCA in a dose-dependent manner (p < 0.01) but did not significantly reduce NCA induced by leukotriene B(4) or complement-activated serum. The reducing agents, dithionite, deferoxamine, and dithiothreitol, reversed and exogenous L-tyrosine abrogated the peroxynitrite-induced NCA inhibition. Papa-NONOate [N-(3-ammoniopropyl)-N-(n-propyl)amino]diazen-1-ium-1, 2-dialase or sodium nitroprusside, NO donors, or a combination of xanthine and xanthine oxidase to generate superoxide did not show an inhibitory effect on NCA induced by IL-8. In contrast, small amounts of SIN-1, a peroxynitrite generator, caused a concentration-dependent inhibition of NCA by IL-8. Consistent with its capacity to reduce NCA, peroxynitrite treatment reduced IL-8 binding to neutrophils. Nitrotyrosine was detected in the IL-8 incubated with peroxynitrite by enzyme-linked immunosorbent assay. These findings are consistent with nitration of tyrosine by peroxynitrite with subsequent inhibition of IL-8 binding to neutrophils and a reduction in NCA and suggest that oxidants may play an important role in regulation of IL-8-induced neutrophil chemotaxis.  相似文献   

10.
The distribution of the lizard Lacerta schreiberi is likely to have been severely affected by the climatic cycles that have influenced the Iberian Peninsula. Information about the species ecology and Iberian physiogeography was used to generate specific hypotheses about episodes of colonization and subsequent population persistence. These hypotheses generated predictions about the distribution of genetic variation, which were tested using nested clade analysis (NCA) supplemented by analysis of molecular variance (amova). Two predictions were confirmed by NCA; that is those that specified multiple and allopatric refugia. However, the remaining three predictions were not corroborated by the analyses. Firstly, a simple analysis of the distribution of genetic variability failed to detect an expected difference in the pattern of colonization between the inland mountain system and the coastal region. Moreover, while NCA did detect the expected genetic pattern in southern coastal populations, it was explained in terms of long-distance migration, which seems implausible because of the extent of unsuitable habitat. A more likely cause of the pattern is population fragmentation and a reduction in population size caused during the Holocene. Finally, NCA also failed to detect a northwestern population expansion, which is supported by other evidence. We conclude that NCA has a limited ability to detect range expansion led by individuals with more ancestral (interior) haplotypes.  相似文献   

11.
Human granulocytes express several species of nonspecific cross-reacting antigens (NCA), glycoproteins belonging to the carcinoembryonic antigen (CEA) family. Our previous studies have shown that at least two different NCA of 95 and 90 kDa are contained in the major NCA band of 90 to 100 kDa detectable upon gel electrophoresis of immunoprecipitates obtained from the cell surfaces of granulocytes with polyclonal anti-NCA. In the present study, the 90 to 100-kDa NCA band was found to include one more species of 100 kDa. This component was reactive with an anti-CD67 antibody as well as polyclonal anti-NCA and released from the cell surface with phosphatidylinositol-specific phospholipase C, indicating that the 100-kDa NCA species is CD67. Both antibodies revealed high binding activities with a recombinant protein of CGM6, which has been identified in a leukocyte cDNA library as an NCA gene and found to encode a glycosyl-phosphatidylinositol-anchored heterotypic cell adhesion molecule. Furthermore, the apparent molecular mass of the deglycosylated CD67 (38 kDa) corresponded with that of the CGM6 protein. These results suggest that CD67 is equivalent to the NCA species CGM6.  相似文献   

12.
原发性肝癌(PLC)患者在精确放疗后乙型肝炎病毒(HBV)再激活是一种常见并发症,及时的预测防护能降低发病率、死亡率。研究表明:多余的特征变量会影响HBV再激活的预测精度。通过提出基于近邻成分分析(NCA)的特征选择方法找出HBV再激活的危险因素及特征组合。之后分别建立经Bayes优化前后的支持向量机模型(SVM)对这些关键特征子集及初始特征集进行分类预测。实验结果表:明HBV DNA水平、KPS评分、分割方式、外放边界、V25、肿瘤分期TNM、ChildPugh等都是影响HBV再激活的危险因素。其中经NCA特征选择之后发现的V25是在乙型肝炎病毒再激活研究中首次提出的危险因素。10折交叉验证下特征组合HBV DNA水平、外放边界、V25的预测精度高达86.11%。支持向量机分类器可以很好的应用于乙型肝炎病毒再激活的研究,特征选择后的关键特征组合具有更优越的分类性能。  相似文献   

13.
In order to investigate the effect of the chiral penultimate unit on the stereoselection of α-amino acid N-carboxyanhydride (NCA) by the terminal unit of a growing chain in the nucleophilic addition-type polymerization, the diastereomers of dipeptide amines, H-(R)-Phe-(S)-Phe-Mo and H-(S)-Phe-(S)-Phe-Mo, in which Mo represents a morpholine residue, were synthesized, and the stereoselectivity in their nucleophilic addition reactions to NCA was investigated and compared with that of a monopeptide amine H-(S)-Phe-OEt. In the reaction with Phe NCA in nitrobenzene, either of the dipeptide amines reacted preferentially with an enantiomer of NCAs having a configuration opposite to the N-terminal unit of the dipeptide amine. The preference of enantiomeric NCA and the extent of stereoselectivity were nearly the same as those found with H-(S)-PheOEt. The opposite-enantiomer selectivity of the dipeptide amines was also observed in the reaction with N-MePhe NCA, and the extent of stereoselectivity was found to increase very much in the reaction of H-(R)-PHe-(S)-Phe-Mo compared with that of H-(S)-Phe-OEt. Therefore, the enhancement of the stereoselectivity of the N-terminal unit by the penultimate unit was shown experimentally. On the other hand, the stereoselectivity of H-(S)-Phe-(S)-Phe-Mo was not very different from that of H-(S)-Phe-OEt. These results were obtained either in nitrobenze or in m-dimethoxybenzene. H-(S)-Phe-(S)-Phe-OEt tends to aggregate by an intermolecular hydrogen bond in aqueous and tetrahydrofuran solutions. Its pKa value and nucleophilicity towards NCA were much lower than H-(R)-Phe-(S)-Phe-Mo, which was free from the aggregation under similar conditions. These experimental results suggest that the major product in the polymerization of (RS)-Phe NCA by amine should be an alternating copolymer. However, this prediction was not verified experimentally, and the important contributions from the aggregation and the molecular weight distribution of growing chains were suggested.  相似文献   

14.
Catalases are key regulators of reactive oxygen species homeostasis in plant cells. However, the regulation of catalase activity is not well understood. In this study, we isolated an Arabidopsis thaliana mutant, no catalase activity1-3 (nca1-3) that is hypersensitive to many abiotic stress treatments. The mutated gene was identified by map-based cloning as NCA1, which encodes a protein containing an N-terminal RING-finger domain and a C-terminal tetratricopeptide repeat-like helical domain. NCA1 interacts with and increases catalase activity maximally in a 240-kD complex in planta. In vitro, NCA1 interacts with CATALASE2 (CAT2) in a 1:1 molar ratio, and the NCA1 C terminus is essential for this interaction. CAT2 activity increased 10-fold in the presence of NCA1, and zinc ion binding of the NCA1 N terminus is required for this increase. NCA1 has chaperone protein activity that may maintain the folding of catalase in a functional state. NCA1 is a cytosol-located protein. Expression of NCA1 in the mitochondrion of the nca1-3 mutant does not rescue the abiotic stress phenotypes of the mutant, while expression in the cytosol or peroxisome does. Our results suggest that NCA1 is essential for catalase activity.  相似文献   

15.
The polymerization of DL -β-phenylalanine N-carboxyanhydride (NCA) initiated by poly(N-benzylglycine)diethylamide (DEA) and poly(N-methyl-DL -alanine)DEA has been investigated. As previously reported, polysarcosine DEA, poly-N-ethylglycine DEA, and poly-N-n-propylglycine DEA showed marked accelerations in the polymerization of DL -β-phenylalanine NCA as compared with the polymerization initiated by low molecular weight, amines having similar base strength. However, this phenomenon (the chain effect) was not observed with the two polymer catalysts studied in the present investigation With poly-N-methyl-DL -alanine DEA, adsorption of DL -β-phenylalanine NCA onto the polymer chain takes place, though not so effectively as with other polypeptides, so the absence of chain effect was ascribed to a reduced flexibility of the polymer chain. With poly(N-benzylglycine)DEA, the reactivity of terminal base group was found to be much lower than that of other polymer catalysts. However, the absence of the chain effect would be attributed to the rigidity of polymer chain of poly-N-benzylglycine DEA due to the bulkiness of the N-benzyl group.  相似文献   

16.
Retrocerebral glandular complexes of Teleogryllus commodus (Orthoptera : Gryllidae) and Periplaneta americana (Dictyoptera : Blattidae) were examined for 5-hydroxytryptamine (serotonin)-immunoreactive (5-HTi) neurons. In Teleogryllus, a prominent tract of 5-HTi axons crosses the ventral surface of the corpus allatum (CA) from nervus corporis allati 1 (NCA 1), and seems to end at varicosities in NCA 2. Serotoninergic axons within this tract pass cephalad to the corpus cardiacum (CC), which also contains numerous, fine 5-HTi branches. 5-HTi axons originate anteriorly, presumably from the pars intercerebralis (PI) and pars lateralis (PL) of the brain. This is suggested by absence of immunoreactivity at the NCA 2-subesophageal ganglion junction, by intense immunofluorescence of the nervi corporis cardiaci (NCC) 1 and 2, by the presence of 5-HTi perikarya in PI and PL, and by previous data obtained by backfilling NCA 1 and 2. In Periplaneta, 5-HTi varicosities are rare in the CA, but abound in the NCA 2, and in NCC 1, 2, and 3. A few 5-HTi fibers project anteriorly from NCA 2 into the cap-like junction of CA and CC, and some traverse the CA to enter the postallatal nerves. Large, 5-HTi axons of NCC 3 ramify within the CC, while others contribute to an anterior branch of NCA 2. As in Teleogryllus, it is unlikely that 5-HTi fibers in NCA 2 originate from somata in the subesophageal ganglion. When cobalt staining and serotonin immunocytochemistry were combined to stain subesophageal neurons of Periplaneta, 5-HTi somata could not be paired with those back-filled via NCA 2. Conspicuous 5-HTi tracts between NCA 2 and the CC of Teleogryllus have no counterpart in Periplaneta.  相似文献   

17.
Antigenic variants of the nonspecific cross-reacting antigen (NCA)   总被引:2,自引:0,他引:2  
Monoclonal antibodies (Mab) were prepared against nonspecific cross-reacting antigen (NCA) and were selected on the basis of their absence of reactivity with carcinoembryonic antigen (CEA). Four Mab were found which allowed the characterization on CEA of three epitopes, defined A, B, and C. These epitopes were all located on the peptidic moiety of this highly glycosylated antigen and were present on NCA molecules of heterogeneous m.w. (greater than 100,000, 80,000, and 48,000 m.w., the latter being the most abundant). The amount of NCA was estimated in 251 human sera both by a conventional RIA, using a rabbit antiserum, and by EIA, using different Mab: Mab 4, 18, and 33, which reacted, respectively, with epitopes A, B, and C. Each assay gave a different value of the absolute concentration of NCA in the serum. On the whole, Mab 4 gave lower values, whereas Mab 18 and 33 gave higher values as compared to RIA. Furthermore, whereas all of the human sera contained NCA which was measurable by RIA, 67 sera typed negative in EIA when using Mab 4 or 18. Eight additional sera were negative in more than one EIA. Negativity when using Mab 33 was observed in only one serum, which was also negative with Mab 4 and 18. Twenty-five of 30 sera which were negative with Mab 4 came from cancer patients, and 32 of 37 sera negative with Mab 18 came from normal subjects and noncancer patients, giving a statistically highly significant difference between the two groups of sera (p less than 0.001). Analysis of tissue perchloric extracts and NCA samples purified from these extracts gave similar results. Three extracts (one from lung, two from cancer tissue) and the corresponding NCA samples were negative with Mab 18. The discrepancies observed in these assays are best explained by assuming the existence of antigenic variants of NCA which have not been described previously. These variants appear to exist in various proportions in the different sera. The variants may represent antigenically complete and incomplete molecules. Alternatively, most of the NCA molecules may be incomplete, lacking one or another of the several NCA-specific epitopes. Sequential immunoprecipitation experiments were in favor of the second hypothesis, showing that most of the NCA molecules were incomplete, lacking either epitope A or B.  相似文献   

18.
Polymerizations of L - and DL -phenylalanine N-carboxyanhydride in nitrobenzene by poly (N-methyl-L -alanine) of varying degrees of polymerization (n = 1–30) were investigated. Poly(N-methyl-L -alanine) was prepared by the polymerization of N-methyl-L -alanine NCA with N-methyl-L -alanine diethylamide and the degree of polymerization was controlled by the molar ratio [NCA]/[Catalyst] + 1. This polymer was shown to be an asymmetrically selective catalyst which polymerized L -phenylalanine NCA at a faster rate than DL -phenylalanine NCA. With increasing degree of polymerization the stability of the secondary structure of poly(N-methyl-L -alanine) increased. This was confirmed by circular dichroism spectra. However, the degree of asymmetric selection did not increase as the stability of the secondary structure of poly(N-methyl-L -alanine) increased. These findings indicate that the interaction of a growing polypeptide in an ordered structure with NCA molecules prior to the reaction does not lead to an asymmetric selection, and that the mechanism of the asymmetric selection by poly(N-methyl-L -alanine) should be different from those proposed so far.  相似文献   

19.
Severance of nervi corporis allati I (NCA I) in day-1 adult female Locusta migratoria resulted in a significant decrease and a loss of the characteristic pattern of juvenile hormone biosynthesis by the corpora allata as determined by radiochemical assay. This decrease in the rate of juvenile hormone biosynthesis was not reflected in basal oöcyte growth. The lengths of the oöcytes were the same in NCA-transectioned and in the sham-operated females. The effect of severance of both NCA I and NCA II on juvenile hormone biosynthesis and ovarian maturation was similar to the effect of NCA I severance only.Rate of juvenile hormone biosynthesis by corpora allata of fourth-instar larvae exhibited a maximum of activity in the middle of the stadium. The severance of NCA I early in the stadium resulted in a very low rate of juvenile hormone biosynthesis and a disappearance of this peak. In NCA I-transectioned larvae, the duration of the stadium was significantly increased although larvae moulted into normal fifth instar.  相似文献   

20.
A cardiaca-allatal commissural plexus (CACP) lies between and partly overlapping the postcommissural lobes of the corpora cardiaca (CC), the nervi corpori allati I (NCA I) and the corpora allata (CA). CACP, which is often continuous posteriorly with a complicated postallatal plexus (PAP), comprises a variable number of connectives with neurosecretory processes linking the cardiaca-commissural organ or dorsal cardiac commissure (containig tritocerebral fibres) to the NCA I. the allatal commissure and the CA. Neurosecretory processes are exchanged between the two halves of the cephalic neuroendocrine complex (CNC) both intracerebrally at different locales, possibly to ensure functional synchrony of CNC components. NCA I and CACP are drawn out with their stroma to varying extents over the CA. Histophysiological evidence suggests that part of the stainable secretion stored in, and or in axonal transit through CA may be released through CA surface; NCA I, the nervi cardiostomatogastrici, CACP, perhaps also NCA II may function as neurohaemal areas. A “directed” neurosecretory pathway could be distinguished from PAP to the foregut and the fat body. The degree of spatial intimacy detected between neurosecretory and stomatogastric components of CNC suggests that the two systems may function in an integrated fashion. The recurrent-oesophageal nerve complex serves not only for a direct transport of neurosecretion, but also as one of the sites of its release.  相似文献   

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