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1.
Glycogen debranching enzyme: purification, antibody characterization, and immunoblot analyses of type III glycogen storage disease. 总被引:2,自引:1,他引:1 下载免费PDF全文
Type III glycogen storage disease is caused by a deficiency of glycogen debranching-enzyme activity. Many patients with this disease have both liver and muscle involvement, whereas others have only liver involvement without clinical or laboratory evidence of myopathy. To improve our understanding of the molecular basis of the disease, debranching enzyme was purified 238-fold from porcine skeletal muscle. In sodium dodecyl sulfate-polyacrylamide gel electrophoresis the purified enzyme gave a single band with a relative molecular weight of 160,000 that migrated to the same position as purified rabbit-muscle debranching enzyme. Antiserum against porcine debranching enzyme was prepared in rabbit. The antiserum reacted against porcine debranching enzyme with a single precipitin line and demonstrated a reaction having complete identity to those of both the enzyme present in crude muscle and the enzyme present in liver extracts. Incubation of antiserum with purified porcine debranching enzyme inhibited almost all enzyme activity, whereas such treatment with preimmune serum had little effect. The antiserum also inhibited debranching-enzyme activity in crude liver extracts from both pigs and humans to the same extent as was observed in muscle. Immunoblot analysis probed with anti-porcine-muscle debranching-enzyme antiserum showed that the antiserum can detect debranching enzyme in both human muscle and human liver. The bands detected in human samples by the antiserum were the same size as the one detected in porcine muscle. Five patients with Type III and six patients with other types of glycogen storage disease were subjected to immunoblot analysis. Although anti-porcine antiserum detected specific bands in all liver and muscle samples from patients with other types of glycogen storage disease (Types I, II, and IX), the antiserum detected no cross-reactive material in any of the liver or muscle samples from patients with Type III glycogen storage disease. These data indicate (1) immunochemical similarity of debranching enzyme in liver and muscle and (2) that deficiency of debranching-enzyme activity in Type III glycogen storage disease is due to absence of debrancher protein in the patients that we studied. 相似文献
2.
Zenghui Hu Yingbai Shen Fanyi Shen Youqing Luo Xiaohua Su 《Trees - Structure and Function》2009,23(5):1003-1011
Interplant communication has been widely demonstrated in plants, especially in herbaceous plants. In this study, mechanical
damage was shown to affect the levels of pyrochatechol, chlorogenic acid, gallic acid and p-hydroxyl benzoic acid in poplar (Populus simonii × P. pyramidalis ‘Opera 8277’) cuttings, indicating the activation of defense response. In neighboring intact cuttings, the levels of those
phenolics also varied when compared to the control, suggesting the interplant communication between poplar cuttings. Three
volatiles, methyl jasmonate, methyl salicylate and benzothiazole, were detected in volatiles emitted from mechanically damaged
poplar cuttings. All of them can induce changes in the levels of four phenolics. Therefore, they could act as airborne signals
between P. simonii × P. pyramidalis ‘Opera 8277’ cuttings. The different change patterns of phenolic contents induced by different volatiles imply that the defense
response activated in neighboring plants may be regulated by multiple signaling pathways. The results also suggest that the
entire defense response of plants involves a variety of airborne signals in wound-induced volatiles. 相似文献
3.
The UDS induced in cultured FL cells by exposure to chemicals was measured as hydroxyurea-resistant incorporation of 3H-TdR in the acid-insoluble fraction of the 14C-TdR-prelabelled cells synchronized by the combination of arginine starvation and pretreatment with hydroxyurea. The level of UDS is represented by the ratios of 3H/14C radioactivities which are measures of specific activities of 3H. Two direct-acting alkylating agents, MMS and MNNG, a cross-linking agent, mitomycin C, and 3 procarcinogens, B(a)P, AFB1 and cyclophosphamide elicited UDS in the absence or presence of the liver-metabolizing system. Three chemicals of unknown carcinogenicity were also able to induce UDS in this assay system, i.e., bis-(O,O-diethylphosphinothioyl)-disulphide, 4-chlorophenoxy acetic acid (sodium salt) and caramelized malt sugar. With the exception of 4-chlorophenoxy acetic acid, they were also active in the Ames test. 相似文献
4.
The mass transfer in a hemodialyser in the presence of combined dialysis and ultrafiltration has been calculated by integration of mass fluxes across the boundary layers in blood and dialysate phase taking into account the partial rejection of solute as well as changes in local blood flow and ultrafiltration flux along the membrane. Clearances of creatinin, vitamin B12, and myoglobin have been calculated as a function of blood and ultrafiltrate flow rate and were found to be in good agreement with in vitro measurements. The data suggest the following empirical correlation for the hemodiafiltration clearance. 相似文献
5.
6.
利用蛋白质内源荧光和酶活性两种信号以及荧光偏振,HPLC和停流等方法研究了盐酸胍诱导的α淀粉酶去折叠与重折叠的平衡转变和动力学。实验结果表明α淀粉酶去折叠与重折叠是两个不同的过程;变性与复性过程中可能伴有聚集体生成;去折叠与重折叠均为双相过程,重折叠大约始于2秒之后。 相似文献
7.
蛋白质结构成对比较的新方法 总被引:7,自引:3,他引:4
介绍一种蛋白质三维结构的快速比较方法.此方法毋需初始联配,而能自动寻找和智能迭代.利用本程序对珠蛋白、丝氨酸蛋白酶、天冬氨酸蛋白酶、钙结合蛋白和溶菌酶作了系统的结构比较,取得了满意的结果.本文也讨论了衡量联配结果好坏的要素问题. 相似文献
8.
中药固真方对一些与细胞增殖有关基因表达的影响姚明忠,顾文聪,丁卫,韩志芬,杜国光(上海中医药大学生物化学教研室,上海200032)(北京医科大学生物化学与分子生物学系,北京100083)中药固真方(VRF)具有补肾益精、延缓衰老的作用[1].能提高成... 相似文献
9.
用σ ̄(70)或σ ̄(38)和核心酶(E)组成的大肠杆菌RNA聚合酶全酶(Eσ)对四种启动子进行了体外转录。结果表明:lacUV5,rplJ主要被Eσ ̄(70)识别,katE主要被Eσ ̄(38)识别,fic在低盐浓度时被Eσ ̄(70)识别,在高浓度盐时被Eσ ̄(38)识别;Eσ ̄(38)转录特异性启动子所需酶量大于Eσ ̄(70)转录特异性启动子的酶量;在含有分别对σ ̄(70)或σ ̄(38)亲和性大的混和启动子的体外转录中,启动子之间不存在干扰和竞争,转录水平与启动子浓度成正相关。 相似文献
10.
罗汉果双受精过程的细胞学观察 总被引:3,自引:1,他引:2
罗汉果(Siraitiagrosvenori(Swingle)C.Jemey)双受精过程属有丝分裂前配子融合类型,授粉后24~48h,花粉管进入胚囊,穿过一个助细胞,放出两个精子。雌雄核融合和雄核与次生核融合同时发生在授粉后62~72,雄核与次生核融合速度快于配子融合,72h后即可见到初生胚乳核分裂。合子中的雌雄核仁在授粉后第5~6d融合,授粉后8~9d合成分裂形成二细胞胚。在双受精过程中,多次观察到有多条花粉管进入胚囊和多精入极核现象。原胚期有附加花粉管从珠孔进入。 相似文献