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51.
Isolation and RNA-binding analysis of NAD+-isocitrate dehydrogenases from Kluyveromyces lactis and Schizosaccharomyces pombe 总被引:1,自引:0,他引:1
Krebs cycle NAD+-isocitrate dehydrogenase (Idh) binds to the 5-UTRs of all mitochondrial mRNAs in Saccharomyces cerevisiae. We hypothesize that this leader-binding activity plays a role in translational regulation, thereby linking mitochondrial
biogenesis to the need for respiratory function. Analysis of effects of leader binding on mitochondrial translation is complicated
by the involvement of the enzyme in mitochondrial metabolism. We have therefore searched for an Idh altered in RNA binding,
but retaining full enzyme activity. Idh from Kluyveromyces lactis and Schizosaccharomyces pombe was partially purified and examined for the ability to bind Cox2 mRNA. Sch. pombe Idh, like the S. cerevisiae enzyme, has high affinity for both its own, K. lactis and S. cerevisiaeCOX2 leaders. In contrast, Idh purified from K. lactis shows only low affinity for all mRNAs tested. To determine what distinguishes K. lactis Idh from S. cerevisiae Idh, genes encoding the two subunits of Idh in K. lactis were cloned and sequenced. Sequence comparison revealed high levels of similarity throughout the proteins, in particular
in regions involved in enzyme activity, co-factor and regulator binding. Non-conserved residues between the subunits from
the two yeasts are candidates for involvement in the interaction with RNA.
Received: 19 January 2000 / 24 March 2000 相似文献
52.
N. Zurgil R. Bakimer M. Kaplan P. Youinou Y. Shoenfeld 《Journal of clinical immunology》1991,11(5):239-245
Antimitochondrial antibodies (AMA) may be detected in 95% of patients with primary biliary cirrhosis (PBC). The target autoantigens for the AMA were recently identified as four closely related metabolic enzymes located in the mitochondria. We have purified the pyruvate dehydrogenase (PDH) enzyme from bovine heart, showing that all PBC sera reacted with a 74-kd band. PDH was utilized to establish an ELISA assay for detecting the relevant antibodies. One hundred twelve of 120 sera from patients with PBC (95%) reacted with the PDH but none of the 201 control sera, including normal subjects and a panel of sera from other patients with liver diseases, showed similar reactivity. In 77% of the PBC sera the anti-PDH antibody isotype was identified as a combination of IgG and IgM, while in 18% only IgM was detected. In 5% of the sera the isotype was confined to IgG. PBC sera specifically inhibited the PDH enzyme activity. The enzyme inhibition correlated with the anti-PDH antibody titers. Thus, PDH seems to be one of the major target epitopes for AMA observed in sera of patients with PBC. 相似文献
53.
L. B. Kim 《Bulletin of experimental biology and medicine》2008,146(6):680-681
We studied enzyme systems (lactate dehydrogenase) of mitochondria in cerebral nerve cells in experimental encephalopathy developing
after thermal injury. In animals receiving neuromedin at the early terms after injury, the ratio of forward to reverse lactate
dehydrogenase reactions significantly increased over the first day after injury and returned to normal on day 7.
__________
Translated from Byulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 145, No. 6, pp. 626–627, June, 2008 相似文献
54.
55.
The objective was to investigate glucose-6-phosphate dehydrogenase (G6PD) activity in monolayer cultures of thyroid epithelial cells and to examine whether inhibition of nitric oxide synthase affects activity of G6PD or oxygen sensitivity of the assay. Primary cultures without TSH addition prior to experiments demonstrated a TSH-dependent increase in G6PD activity. G6PD activity was higher in F12 medium than in a serum-free physiological medium. Secondary cultures grown in F12 medium demonstrated a diminished activity of G6PD and a lack of response to TSH. In the serum-free physiological medium, G6PD activity was comparable to that found in primary cultures and a response to high concentrations of TSH was maintained. In primary cultures grown in F12 medium devoid of TSH, G6PD activity decreased dose-dependently when nitric oxide synthase activity was inhibited. The oxygen sensitivity of the assay was comparable to that reported previously in malignant cells and correlated with the activity of G6PD in primary cultures. We suggest that thyroid epithelial cells may be an appropriate system to investigate oxygen sensitivity of the G6PD assay as the cells demonstrate a reduced oxygen sensitivity which can be influenced by culture conditions. 相似文献
56.
57.
阿霉素性心肌病之心肌线粒体酶的细胞化学研究 总被引:2,自引:0,他引:2
阿霉素性心肌病之心肌线粒体SDH和CCO的细胞化学观察发现,多数线粒体的酶活性较弱,少数较强或中等,并与ADR的累积用量和心肌损伤程度有关。作者认为SDH和CCO活性降低,线粒体能量代谢障碍在ADR引起心肌病的过程中起着重要作用。 相似文献
58.
ADA、ACE、LDH、CEA的联合检测在结核性和恶性胸腔积液中的鉴别诊断价值 总被引:1,自引:0,他引:1
目的测定血清和胸水中腺苷脱氨酶(ADA)、血管紧张素转化酶(ACE)、乳酸脱氢酶(LDH)与癌胚抗原(CEA)的水平,探讨其指标联合检测对结核性和恶性胸水的鉴别诊断意义。方法对临床已确诊的72例胸腔积液患者(结核性40例,恶性32例)的胸水和血清分别采用酶免疫法和化学发光法进行ADA、ACE、LDH和CEA含量测定。结果结核性胸水中ADA的含量为(60.2±20.10)U/L,ACE的含量为(35±9.6)U/L,LDH的含量为(338±41)U/L,CEA的含量为(12.8±5.82)μg/L;在恶性胸水中,ADA为(11.02±5.23)U/L,ACE为(16±11.0)U/L,LDH为(379±69.0)U/L,CEA为(39.9±19.7)μg/L。结核性胸水ADA和ACE含量较恶性胸水组明显增高(P〈0.01),CEA在恶性胸水中含量较结核性胸水组明显增高(P〈0.01)。胸水中ADA和ACE的检测对结性性胸膜积液诊断的敏感性分别为84.3%、87.5%,特异性分别为87.5%、80.0%;而胸水中LDH和CEA的检测对恶性胸膜积液诊断的敏感分别为84.3%、75.0%,特异性分别为80.0%、93.0%。四项指标联合检测敏感性性为78.1%,特异性为97.5%,较单一指标的特异性高。结论胸水中ADA、ACE、LDH和CEA的联合检测对结核性和恶性胸水的鉴别诊断具有一定价值,有助于临床胸水性质的诊断。 相似文献
59.
Micro-organisms have developed systems to adapt to sudden changes in the environment. Here we describe the response of the yeastSaccharomyces cerevisiae to osmotic stress. A drop in the water activity (aw) of the medium following the addition of NaCl led to an immediate shrinkage of the cells. During the 2 h following the osmotic shock the cells partially restored their cell volume. This process depended on active protein synthesis. During the recovery period the cells accumulated glycerol intracellularly as a compatible solute and very little glycerol was leaking out of the cell. We have investigated in more detail the enzymes of glycerol metabolism and found that only the cytoplasmic glycerol-3-phosphate dehydrogenase was strongly induced. The level of induction was dependent on the yeast strain used and the degree of osmotic stress. The synthesis of cytoplasmic glycerol-3-phosphate dehydrogenase is also regulated by glucose repression. Using mutants defective in glucose repression (hxk2), or derepression (snf1), and with invertase as a marker enzyme, we show that glucose repression and the osmotic-stress response system regulate glycerol-3-phosphate dehydrogenase synthesis independently. We infer that specific control mechanisms sense the osmotic situation of the cell and induce responses such as the production and retention of glycerol. 相似文献
60.
Post-exercise ketosis and the glycogen content of liver and muscle in rats on a high carbohydrate diet 总被引:1,自引:0,他引:1
Summary Post-exercise ketosis is known to be suppressed by physical training and by a high carbohydrate diet. As a result it has often
been presumed, but not proven, that the development of post-exercise ketosis is closely related to the glycogen content of
the liver. We therefore studied the effect of 1 h of treadmill running on the blood 3-hydroxybutyrate and liver and muscle
glycogen concentrations of carbohydrate-loaded trained (n=72) and untrained rats (n=72). Resting liver and muscle glycogen levels were 25%–30% higher in the trained than in the untrained animals. The resting
3-hydroxybutyrate concentrations of both groups of rats were very low: <0.08 mmol·1−1. Exercise did not significantly influence the blood 3-hydroxybutyrate concentrations of trained rats, but caused a marked
post-exercise ketosis (1.40±0.40 mmol·1−1 1 h after exercise) in the untrained animals, the time-course of which was the approximate inverse of the changes in liver
glycogen concentration. Interpreting the results in the light of similar data obtained after a normal and low carbohydrate
diet it has been concluded that trained animals probably owe their relative resistance to post-exercise ketosis to their higher
liver glycogen concentrations as well as to greater peripheral stores of mobilizable carbohydrate. 相似文献