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91.
Novel yeast protein kinase (YPK1 gene product) is a 40-kilodalton phosphotyrosyl protein associated with protein-tyrosine kinase activity. 总被引:23,自引:8,他引:15
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D Dailey G L Schieven M Y Lim H Marquardt T Gilmore J Thorner G S Martin 《Molecular and cellular biology》1990,10(12):6244-6256
Extracts of bakers' yeast (Saccharomyces cerevisiae) contain protein-tyrosine kinase activity that can be detected with a synthetic Glu-Tyr copolymer as substrate (G. Schieven, J. Thorner, and G.S. Martin, Science 231:390-393, 1986). By using this assay in conjunction with ion-exchange and affinity chromatography, a soluble tyrosine kinase activity was purified over 8,000-fold from yeast extracts. The purified activity did not utilize typical substrates for mammalian protein-tyrosine kinases (enolase, casein, and histones). The level of tyrosine kinase activity at all steps of each preparation correlated with the content of a 40-kDa protein (p40). Upon incubation of the most highly purified fractions with Mn-ATP or Mg-ATP, p40 was the only protein phosphorylated on tyrosine. Immunoblotting of purified p40 or total yeast extracts with antiphosphotyrosine antibodies and phosphoamino acid analysis of 32P-labeled yeast proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis indicated that the 40-kDa protein is normally phosphorylated at tyrosine in vivo. 32P-labeled p40 immunoprecipitated from extracts of metabolically labeled cells by affinity-purified anti-p40 antibodies contained both phosphoserine and phosphotyrosine. The gene encoding p40 (YPK1) was cloned from a yeast genomic library by using oligonucleotide probes designed on the basis of the sequence of purified peptides. As deduced from the nucleotide sequence of YPK1, p40 is homologous to known protein kinases, with features that resemble known protein-serine kinases more than known protein-tyrosine kinases. Thus, p40 is a protein kinase which is phosphorylated in vivo and in vitro at both tyrosine and serine residues; it may be a novel type of autophosphorylating tyrosine kinase, a bifunctional (serine/tyrosine-specific) protein kinase, or a serine kinase that is a substrate for an associated tyrosine kinase. 相似文献
92.
Decarboxylation of uroporphyrinogen III by erythrocyte uroporphyrinogen decarboxylase. Evidence for a random decarboxylation mechanism.
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The isomeric composition of type-III heptacarboxylic porphyrinogens derived from decarbosylation of uroporphyrinogen III by erythrocyte uroporphyringogen decarboxylase was analysed by h.p.l.c. with electrochemical detection. All four possible isomers were identified, and there were little differences in the proportion of isomers formed by erythrocytes from normal subjects and from patients with sporadic porphyria cutanea tarda. The results provide conclusive evidence that the normal decarboxylation pathway is random in nature, and the fourth isomer only increases when enzyme abnormality is found. 相似文献
93.
Wei de Zhang Fan Qing Li Xiao Yun Zhang Ying Chen 《Biological trace element research》1988,17(1):81-90
The growth of the protozoanBlepherisma is stimulated by Lanthanum (La) at concentrations as low as 0.32 ppm. In mice Yttrium (Y) and Ytterbium (Yb) are absorbed,
accumulated, and metabolized. Both rare earth elements (RE) exhibit a high affinity for teeth and bones, accumulation occurs
and metabolism is slow. In the livers of RE-exposed mice, concentrations are variable. The liver is apparently capable of
absorbing and discharging RE in a manner depending on metabolic activity. The main route of discharge for ingested REs is
the alimentary canal. Exposure of pregnant mice to RE leads to rapid placental transfer of RE; 14.1% of the total amount of
RE administered was detected in newborn mice. Young, developing organisms appear to be especially susceptible to RE accumulation. 相似文献
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八名射击运动员(手枪2人,步枪6人),在实弹射击训练中,连续同步地记录了脑电、心电、心率、阻抗呼吸波。同时记录了受试者持枪稳定的程度。 分析表明,射击环数是五项独立无关的指标综合作用的结果。本文首次对射击时生理指标和射击环数进行系统辨识建模分析,求出每个人的系统参数_i值。找出了每人射击时最佳生理参数,为指导射击练习和反馈训练,提供了客观生理依据。 本文设计的激光光点录相方法,可反映持枪稳定的程度,简单、方便,可在射击训练中采用。 相似文献
98.
<正> 甲型血友病是最常见的遗传性凝血疾病,是由于凝血因子Ⅷ(FVⅢ)基因缺陷所致。目前对此病尚无满意的治疗方法。利用DNA分析技术进行甲型血友病基因检测与产前基因诊断对开展优生优育有重要的实际应用价值。由于FⅧ基因组织结构庞大,分子病理改变复杂,目前多采用DNA限制酶片段长度多态性(RFLPs)为遗传标志对有缺陷的FⅧ基因进行连锁分析。St 14(DXS52)是人X染色体长臂远端的一段基因外DNA序列,与FⅧ基因紧密连锁。国外已将St14/Taq Ⅰ RFLPs用于甲型血友病基因携带者的检测,但尚未见到用于产前基因诊断的报道。我们利用引进的St14片段为探针,采用简化的低脂奶粉杂交体系和DNA凝胶原 相似文献
99.
<正> 近年来,光敏亲和标记技术已被广泛应用于激素及共受体,蛋白和核酸的相互作用、酶的结构与功能、膜蛋白结构、tRNA同共合成酶的识别等研究上,鉴于ATP是某些酶的底物,因此合成光敏的ATP底物类似物将有助于国内对达些酶的结构与功能进行深入研究,为此我们合成了8-N_8-5′-ATP,现将实验结果报道如下: 相似文献
100.
测定了3T3细胞、人和大鼠一些组织中DNA拓扑异构酶Ⅰ的活性;估计了核酸内切酶对拓扑酶Ⅰ松弛活性测定的干扰程度;发现增殖组织全细胞抽提液中酶比活高于正常分化组织,而且在异常增殖组织中酶比活的增高更为显著。 相似文献