Structural characterization of zinc-deficient human superoxide dismutase and implications for ALS

Over 130 mutations to copper, zinc superoxide dismutase (SOD) are implicated in the selective death of motor neurons found in 25% of patients with familial amyotrophic lateral sclerosis (ALS). Despite their widespread distribution, ALS mutations appear positioned to cause structural and misfolding defects. Such defects decrease SOD's affinity for zinc, and loss of zinc from SOD is sufficient to induce apoptosis in motor neurons in vitro. To examine the importance of the zinc site in the structure and pathogenesis of human SOD, we determined the 2.0-A-resolution crystal structure of a designed zinc-deficient human SOD, in which two zinc-binding ligands have been mutated to hydrogen-bonding serine residues. This structure revealed a 9 degrees twist of the subunits, which opens the SOD dimer interface and represents the largest intersubunit rotational shift observed for a human SOD variant. Furthermore, the electrostatic loop and zinc-binding subloop were partly disordered, the catalytically important Arg143 was rotated away from the active site, and the normally rigid intramolecular Cys57-Cys146 disulfide bridge assumed two conformations. Together, these changes allow small molecules greater access to the catalytic copper, consistent with the observed increased redox activity of zinc-deficient SOD. Moreover, the dimer interface is weakened and the Cys57-Cys146 disulfide is more labile, as demonstrated by the increased aggregation of zinc-deficient SOD in the presence of a thiol reductant. However, equimolar Cu,Zn SOD rapidly forms heterodimers with zinc-deficient SOD (t1/2 approximately 15 min) and prevents aggregation. The stabilization of zinc-deficient SOD as a heterodimer with Cu,Zn SOD may contribute to the dominant inheritance of ALS mutations. These results have general implications for the importance of framework stability on normal metalloenzyme function and specific implications for the role of zinc ion in the fatal neuropathology associated with SOD mutations.     

Inactivation of metabolic enzymes by photo-treatment with zinc meta N-methylpyridylporphyrin

Cell proliferation is notably dependent on energy supply and generation of reducing equivalents in the form of NADPH for reductive biosynthesis. Blockage of pathways generating energy and reducing equivalents has proved successful for cancer treatment. We have previously reported that isomeric Zn(II) N-methylpyridylporphyrins (ZnTM-2(3,4)-PyP⁴⁺) can act as photosensitizers, preventing cell proliferation and causing cell death in vitro. The present study demonstrates that upon illumination, ZnTM-3-PyP inactivates glucose-6-phosphate dehydrogenase, glyceraldehyde-3-phosphate dehydrogenase, lactate dehydrogenase, NADP⁺ -linked isocitrate dehydrogenase, aconitase, and fumarase in adenocarcinoma LS174T cells. ZnTM-3-PyP⁴⁺ was significantly more effective than hematoporphyrin derivative (HpD) for inactivation of all enzymes, except aconitase and isocitrate dehydrogenase. Enzyme inactivation was accompanied by aggregation, presumably due to protein cross-linking of some of the enzymes tested. Inactivation of metabolic enzymes caused disruption of cancer cells' metabolism and is likely to be one of the major reasons for antiproliferative activity of ZnTM-3-PyP.     

不同季节高原鼢鼠乳酸脱氢酶活力和同工酶谱

目的:探讨高原鼢鼠对洞道低氧高二氧化碳环境的代谢适应机制。方法:用酶活力分析法,分析春季、夏季和秋季高原鼢鼠血清乳酸脱氢酶(LDH)活力、乳酸含量和组织LDH活力,用聚丙烯酰胺凝胶电泳法分析血清和组织LDH同工酶谱。结果:高原鼢鼠血清LDH活力在春夏秋三季具有明显的差异,春季高于夏季,夏季高于秋季,血清乳酸含量表现出同样的变化趋势;春季血清中五种同工酶条带都清晰可见,夏季血清中LDH5和LDH4清晰可见,秋季血清中只能看见LDH5带。骨骼肌、心肌和脑组织LDH活力较高,而且从春季到秋季显著降低;肝、肾和肺组织LDH活力较低,肝组织LDH活力春季显著高于夏季和秋季,夏秋两季之间没有明显差异;肾和肺组织LDH活力在春季与夏季之间没有明显差异,但秋季明显降低。心、肝、肺、肾、脑和肌肉组织LDH同工酶谱,在春夏秋三季都显示出五条带,并表现出明显的组织差异;各组织同工酶含量也有不同程度的季节差异。结论:高原鼢鼠体内糖酵解过程具有明显的季节性变化,从春季到秋季依次降低,这与它们的季节性活动特点和洞道中氧气和二氧化碳的季节性波动有关。     

珍稀濒危植物珙桐超氧化物歧化酶活性

采用比色法测定了不同年龄和部位珙桐叶的超氧化物歧化酶(SOD)活性,以探讨不同年龄级珙桐的生理生态适应性。结果表明:不同年龄同一生长期珙桐叶的SOD活性不同,同年龄级同层珙桐叶的SOD活性随叶位增加大致呈先升后降的变化;同叶位不同层及同层不同叶位的珙桐叶的SOD活性不同。珙桐叶SOD活性对生长期敏感,其活性随叶片的生长、衰老呈先升后降变化。即珙桐叶SOD活性受生物和非生物因子共同影响,且不同年龄级、叶位、层次均存在差异。     

狼毒提取物对枸杞蚜虫的生物活性及其体内酶活性的影响

本文研究了狼毒不同溶剂提取物对枸杞蚜虫的杀虫活性及其体内SOD和GSH—PX活性的影响。结果表明,狼毒的乙醇、三氯甲烷、石油醚和丙酮提取物对枸杞蚜虫具有较强的触杀活性,其中三氯甲烷提取物触杀活性最强;不同提取物触杀处理对枸杞蚜虫体内SOD和GSH—PX具有不同程度的激活作用,活力变化趋势相似,而在内吸处理中,活力变化复杂,4种提取物处理对枸杞蚜虫两种酶系的活性没有明显的抑制作用。     

蛇毒清胶囊对眼镜蛇咬伤局部坏死患者血浆LPO、SOD、GSH-Px的影响

目的观察蛇毒清胶囊对眼镜蛇咬伤合并局部组织坏死患者血浆LPO、SOD、GSH-PX的影响及疗效。方法将眼镜蛇咬伤2 h内合并局部组织坏死的患者60例,随机分为两组,均给予常规治疗,治疗组加服蛇毒清胶囊,均以7天为1疗程。分别于就诊时、咬伤后24 h、疗程结束时测定其血浆LPO、SOD、GSH-PX,比较两组患者三项指标的变化和临床疗效。结果两组患者在就诊时三项指标无明显差异(P>0.05);伤后24 h及疗程结束时,治疗组LPO明显低于对照组,SOD、GSH-PX显著高于对照组,治疗组疗效优于对照组(P<0.05)。结论蛇毒清胶囊能降低眼镜蛇咬伤患者血浆LPO水平,提高其SOD、GSH-PX活性,对眼镜蛇咬伤患者有清除氧自由基的作用。     

Physiological adaptability of three mangrove species to salt stress

The impact of salinity on three arboreal mangrove plants, Sonneratia apetala (Sa), S. caseolaris (Sc) and Rhizophora stylosa (Rs), was studied. The three mangrove species were treated with different salinity levels over a three-month period. The response and adaptation of these three mangrove species to salinity were shown to be different. Net photosynthesis rate, stomata conductance and transpiration rate of leaves decreased and soluble sugar content in leaves increased, with salt concentration in all three mangrove species. The malondial dehyde (MDA) content in stems and leaves of Sa and Sc somewhat decreased when the salinity was lower than 10, but rapidly increased with increasing salt concentration. The MDA content in stems and leaves of Rs increased only when salinity was greater than 40. No changes were observed in the MDA content of roots in the three mangrove species. The adaptabilities of Sa and Sc to salt tolerance were limited. The more salt tolerant the mangrove Rs, the more likely the free oxygen radicals were eliminated through the increase in activity of superoxide dismutase (SOD). Results of this experiment identified salinity levels best suited for the growth and metabolism of the species, which provides information necessary for maintaining mangrove forestation along the South China coast.     

Anti-oxidation and immune responses in mice upon exposure to manganese superoxide dismutase expressed in silkworm larvae, Bombyx mori L

With manganese superoxide dismutase expressed in silkworm larvae, Bomby mori L, we investigate the effects of silkworm larvae powder containing SOD on the antioxidation and the immune system of mouse. The contents of MDA both in mice plasma or liver organ treated with silkworm larvae powder containing manganese superoxide dismutase were reduced compare to control. The superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) activities both in plasma or liver organ of the treated mice were significantly higher than that of both control and bromobenzene treated mice (group-BM), suggesting the silkworm larvae powder containing SOD play a positive role in anti-oxidation in mice. This experiment was also designed to investigate the effects of silkworm larvae powder containing SOD on the immune system of mouse, focused on hemolysin response, hemagglutination against SRBC and the activity of natural killer (NK) cells. All treated mice showed significant increase in hemolysin response to SRBC and demonstrated an activation of NK cell function by the SOD-contained silkworm larvae powder, which suggest a promotion in humoral immunity. The results suggested the SOD expressed in silkworm maybe have potential application in medicine.     

A chromium-tolerant plant growing in Cr-contaminated land

In this paper, a kind of Typhaceae plant species, Typha angustifolia L, with a high tolerance to Cr is described. Experiments were carried out to examine its ability to tolerant Cr and its physiological response. The results showed that there was no difference in growth, plant height, and biomass response to external Cr (VI) between the plants exposed to 100 microM Cr (VI) and control (0 microM), while increasing Cr levels to 200-800 microM induced a significant decrease in plant height and biomass, but no significant injury was detected, even for the plants exposed to 800 microM Cr. Chromium induced significant increases in superoxide dismutase (SOD) and peroxidase (POD) activities. Meanwhile, a significantly positive correlation was found between Cr and Mn or Cu in leaves and roots, respectively. The Cr tolerance of the plant appeared to be associated with the enhancement of SOD and POD activities and the improvement in uptake and translocation of the essential microelements.     

两株内生芽孢杆菌对盐胁迫下大豆幼苗超氧化物歧化酶和过氧化物酶活性影响

【背景】盐胁迫环境严重影响大豆幼苗生长,内生菌可提高作物的抗逆性。【目的】探究接种内生枯草芽孢杆菌127和解蛋白芽孢杆菌133对盐胁迫下大豆幼苗体内超氧化物歧化酶(superoxide dismutase,SOD)和过氧化物酶(peroxidase,POD)活性的影响。【方法】以“徐豆20”为实验材料,采用盆栽实验法,设置对照组、盐胁迫组和盐胁迫接菌组,在人工气候培养条件下,用不同NaCl浓度(50、100、150、200、250和300 mmol/L)处理大豆幼苗,并接种不同OD600值(OD0.33、OD0.50和OD0.75)的菌悬液。【结果】培养14 d,接种枯草芽孢杆菌127的菌悬液OD0.33和OD0.75分别在盐浓度300 mmol/L和100 mmol/L时,SOD活性均为1.04 U/g-FW;接种解蛋白芽孢杆菌133的菌悬液OD0.50在盐浓度300 mmol/L胁迫下POD活性最高为7 820 U/(g·min),对大豆幼苗修复效果较显著。培养28 d,接种枯草芽孢杆菌127的菌悬液OD0.50,在150 mmol/L时SOD活性最高(0.88 U/g-FW);接...