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41.
Distinct tyrosine phosphorylation sites in ZAP-70 mediate activation and negative regulation of antigen receptor function. 总被引:9,自引:4,他引:5
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G Kong M Dalton J B Wardenburg D Straus T Kurosaki A C Chan 《Molecular and cellular biology》1996,16(9):5026-5035
Biochemical and genetic evidence has implicated two families of protein tyrosine kinases (PTKs), the Src- and Syk-PTKs, in T- and B-cell antigen receptor signaling. ZAP-70 is a member of the Syk-PTKs that associates with the T-cell antigen receptor and undergoes tyrosine phosphorylation following receptor activation. Three tyrosine residues, Tyr-292, -492, and -493, have been identified as sites of phosphorylation following T-cell antigen receptor engagement. Utilizing ZAP-70- and Syk-deficient lymphocytes (Syk-DT40 cells), we provide biochemical and functional evidence that heterologous trans-phosphorylation of Tyr-493 by a Src-PTK is required for antigen receptor-mediated activation of both the calcium and ras pathways. In contrast, cells expressing mutations at Tyr-292 or -492 demonstrate hyperactive T- and B-cell antigen receptor phenotypes. Thus, phosphorylation of ZAP-70 mediates both activation and inactivation of antigen receptor signaling. 相似文献
42.
Single-stranded DNA binding protein and DNA helicase of bacteriophage T7 mediate homologous DNA strand exchange. 总被引:7,自引:1,他引:6
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Two proteins encoded by bacteriophage T7, the gene 2.5 single-stranded DNA binding protein and the gene 4 helicase, mediate homologous DNA strand exchange. Gene 2.5 protein stimulates homologous base pairing of two DNA molecules containing complementary single-stranded regions. The formation of a joint molecule consisting of circular, single-stranded M13 DNA, annealed to homologous linear, duplex DNA having 3'- or 5'-single-stranded termini of approximately 100 nucleotides requires stoichiometric amounts of gene 2.5 protein. In the presence of gene 4 helicase, strand transfer proceeds at a rate of > 120 nucleotides/s in a polar 5' to 3' direction with respect to the invading strand, resulting in the production of circular duplex M13 DNA. Strand transfer is coupled to the hydrolysis of a nucleoside 5'-triphosphate. The reaction is dependent on specific interactions between gene 2.5 protein and gene 4 protein. 相似文献
43.
Ejaculates from 3 young boars were collected on 4 occasions as a series of separate 15-ml fractions. The contribution of different fractions of these ejaculates to observed variability in the quality of the semen when used for IVF was then determined. On the basis of sperm concentration, 3 fractions representing the first peak concentration (Fraction 1), the lowest sperm concentration after Fraction 1 (Fraction 2), and the second peak concentration (Fraction 3) were selected for analysis in vitro. Oocyte-cumulus-granulosa cell complexes were obtained by dissection from slaughterhouse ovaries. In vitro matured oocytes were randomly assigned for fertilization by the 3 semen samples from each boar. Sperm concentration was the same in all the samples during prefertilization incubation, while the final concentration for fertilization was 5 x 10(5) sperm/ml. Data were analysed using ANOVA for a split-plot design. In the presence of fraction effects, Student-Newman-Keuls (SNK) test was used for multiple comparison of treatment means. Oocyte penetration rates differed among fractions (P = 0.001) and varied from 69 to 100% (mean 95.7%) for Fraction 1, from 0 to 100% (mean 53.3%) for Fraction 2, and from 50to 100% (mean 89.9%) for Fraction 3. There were also differences in male pronuclear formation rate (P = 0.028; mean 27.6, 9.3 and 16.4% for Fractions 1, 2 and 3, respectively); in the rate of polyspermy (P = 0.0001; mean 92.3, 31.9 and 76.3% for Fractions 1, 2 and 3, respectively); and in the number of penetrated spermatozoa per oocyte P = 0.002; mean 5.58, 1.94 and 4.07 for Fractions 1, 2, and 3, respectively). The first peak concentration of semen (Fraction 1) showed superiority in fertilizing ability and less variability in penetration rate from replicate to replicate compared with the other 2 fractions. By multiple comparison, Boar 1 showed higher rates of penetration (P < 0.05), male pronuclear formation (P < 0.05) and polyspermy (P < 0.05) than the other 2 boars. There was no fraction-by-boar interaction. The IVM-IVF system adopted proved to be a promising method for boar semen evaluation. 相似文献
44.
Ligand binding to a linear lattice composed of N sites, under general conditions of cooperativity and number of sites covered upon binding, m, is approached in terms of the theory of contracted partition functions. The partition function of the system obeys a recursion relation leading to a generating function that provides an exact analytical solution for any case of interest. Site-specific properties of the lattice are derived from simple transformations of the analytical expressions. The McGhee-von Hippel model is obtained as a special case in the limit N --> infinity. The derivation is straightforward and involves no combinatorial arguments. Partition functions and site-specific properties are also derived for the case of non-cooperative binding to a two-dimensional torus of length N, containing s sites in its section for a total of sN sites. The torus provides a relevant model for ligand binding to double-stranded DNA (s = 2) or protein helices (s = 3,4). It is proved that non-cooperative binding to the two-dimensional torus can mimic cooperative binding to a one-dimensional linear lattice when m = s. The dimensional embedding of the lattice and the geometry of interaction of its sites play a crucial role in defining the binding properties of the system accessible to experimental measurements. Hence, caution must be exercised in the interpretation of Scatchard plots in terms of the one-dimensional McGhee-von Hippel model, especially when m < or = 4 and the geometry of the system is clearly two-dimensional. 相似文献
45.
丝孢酵母高甲硫氨酸突变株的选育及营养调控 总被引:4,自引:0,他引:4
以丝孢酵母(Trichosporon Behr)ST851为原始菌株,经紫外线诱变,在含乙硫氨酸的双层平板上筛选到多株抗乙硫氨酸突变株。其中ST851-10株抗乙硫氨酸浓度达到350μg/ml,其菌体蛋白质含量由40.5%提高到44.3%,菌体甲硫氨酸含量由20.45mg/g-DCW增加到29.32mg/g-DCW。在以苹果渣为碳源、尿素为氮源、硫酸镁作硫源的最适培养条件下,固态发酵24h后,蛋白质和甲硫氨酸含量较原始菌株分别提高了15.8%和44.9%。培养基中C/N值低有利于甲硫氨酸的合成,C/N值高则适合于菌体生长。在苹果渣固态发酵过程中,适当补加氮源既有利于菌体生长和甲硫氨酸的合成,又可起到调节培养基pH值的作用。 相似文献
46.
沿海防护林防海煞危害初探 总被引:2,自引:0,他引:2
通过后牧村海岸防护林带附近空气中主要盐离子飘尘的观测分析表明,林带对大气盐尘的吸附和截留作用,无林带地区平均沉降量为4.3μg.m^-3.100m^-1,通过林带时为18.4μm^-3.100m^-1。林带的吸队量与树种、枝叶形状有关,侧柏对Cl^-和Na^+的吸附量分别为6330和3350mg.kg^-1,而黑松为1570和950mg.kg^-1,空气中盐尘受天气条件特别是风速的影响,阴天,风速 相似文献
47.
本文用免疫细胞化学方法调查了生长抑素(SOM)免疫反应神经元在人胎丘脑网状核内的分布。流产的胚胎3例,胎龄分别为18周,23周,32周。意外死亡足月新生儿1例、在18周胚胎的丘脑网状核内可见少数染色较浅的SOM免疫反应阳性神经元,呈圆形。从18周到32周,SOM免疫反应阳性细胞数明显增多,突起更丰富。在足月新生儿,SOM阳性细胞数较32周有所减少。结果表明,SOM阳性神经元存在于人胎丘脑网状核内,并且有一定的发育过程。出现于人脑发育的早期阶段,可能在中枢神经系统的发育过程中起重要的作用。 相似文献
48.
本文就植物血细胞凝集素(PHA)作用的不同时间对小鼠脾脏淋巴细胞γ-谷氨酰基转肽酶(γ-GT)活性的影响进行了观察。结果表明,脾脏淋巴细胞转化率随给予PHA时间的延长而上升,对照组<24h组<48h组<72h组;淋巴细胞γ-GT活性于24h明显强于对照组(P<0.001),随着PHA作用时间的延长其γ-GT活性逐渐下降,即48h组与对照组已无显著差异,而72h组却低于对照组(P<0.001)。可见PHA在小鼠体内对γ-GT活性有很大影响。 相似文献
49.
50.
A HPLC method has been developed to measure phosphatidylcholine (PC) containing reactive carbonyl functions in the sn-acyl residue in order to study processes in which such reactive carbonyls can be formed due to e.g. oxidative fragmentation. The method has been applied to determine PC-bound carbonyls as 2, 4-dinitrophenylhydrazones (DNPH) in plasma of burn patients. Plasma from healthy volunteers served as controls. Additionally, in vitro oxidation experiments (A: plasma, buffer diluted; B: plasma + iron-EDTA complex and C: plasma + iron-EDTA complex + H2O2) have been performed to obtain and to identify 2, 4-dinitrophenylhydrazine derivatizable carbonyl functions in plasma PC. Both, the PC-aldehydes and PC-aldehyde DNPH derivatives were cleavable with phospholipase C. Quantification was based on thin-layer chromatography purified soybean phosphatidylcholine, which was identically oxidized and derivatized as the plasma lipids in vitro. 相似文献