Structural transitions upon ligand binding in a cooperative dimeric hemoglobin

Comparison of the 2.4 angstrom resolution crystal structures of dimeric clam hemoglobin in the deoxygenated and carbon-monoxide liganded states shows how radically different the structural basis for cooperative oxygen binding is from that operative in mammalian hemoglobins. Heme groups are in direct communication across a novel subunit interface formed by the E and F helices. The conformational changes at this interface that accompany ligand binding are more dramatic at a tertiary level but more subtle at a quaternary level than those in mammalian hemoglobins. These findings suggest a cooperative mechanism that links ligation at one subunit with potentiation of affinity at the second subunit.     

Blood function in the hydrothermal vent vestimentiferan tube worm

Extracellular hemoglobin in the whole blood of Riftia pachyptila has a high oxygen affinity (P50 = 1.8 millimeters of mercury at 3 degrees C), a moderate decrease in oxygen affinity at higher temperatures (P50 = 2.7 millimeters of mercury at 14 degrees C), a small effect of carbon dioxide on oxygen affinity (Delta log P50/Delta pH =-0.12), and a high oxygen carrying capacity (up to 11 milliliters of oxygen per 100 milliliters of blood). These characteristics are compatible with the high oxygen demand of chemoautotrophic metabolism in the variable vent environment.     

Hemoglobin Kinetics of the Galapagos Rift Vent Tube Worm Riftia pachyptila Jones (Pogonophora; Vestimentifera)

Kinetics of the reactions of Riftia pachyptila hemoglobin with oxygen were followed spectrophotometrically by stopped-flow and laser flash photolysis techniques. The rate of oxygen dissociation increases eightfold over the range of 5 degrees to 20 degrees C (k = 2.2 sec(-1)at 10 degrees C). Oxygen recombination after flash photolysis was biphasic. The rates of both slow and fast phases of the reaction were independent of temperature from 0 degrees to 20 degrees C(k'fast = 7 x 10(6); k'slow = 1 x 16(6) liter mole (-1) sec(-1)). As the oxygen affinity is relatively temperature independent, analysis in terms of the two-state model of cooperativity requires that the conformational equilibrium constant L decrease by about 50-fold between 3 degrees and 15 degrees C.     

4种壳色菲律宾蛤仔在低氧胁迫下的耐受能力比较研究

菲律宾蛤仔Ruditapes philippinarum对环境中溶氧剧烈的波动有较强的适应能力,为了探讨抗氧化酶和热休克蛋白在蛤蛤恣仔的低氧耐受过程中发挥的作用,以对低氧具有不同耐受能力的4种壳色蛤仔(野生蛤仔、斑马蛤、白蛤、白斑马蛤)为研究对象,比较了在1 mg/L的低氧胁迫条件下其抗氧化酶家族基因成员(TPx、CAT、Mn-SOD、Cu/Zn-SOD)和热休克蛋白家族基因成员(HSP40、HSP75、Vps HSP-1、Vps HSP-2)的表达量变化情况。结果表明:在鳃组织中,野生蛤仔的抗氧化酶基因表现出先升高后恢复的趋势,在胁迫后6 h时表达量达到最高值(P0.05),其他3种壳色蛤仔的抗氧化酶基因表现出先降低后恢复的趋势,在胁迫后6 h时表达量达到最低值(P0.05),斑马蛤的TPx基因表达量在胁迫的各个时间点均显著高于其他3种壳色蛤仔(P0.05);在外套膜组织中,4种壳色蛤仔中的抗氧化酶基因均表现出先升高后恢复的趋势,白蛤TPx基因和白斑马蛤CAT基因表达量在胁迫的各个时间点均显著高于其他3种壳色蛤仔(P0.05);在鳃组织中,斑马蛤和白斑马蛤中热休克蛋白家族基因成员表现出先降低后升高再降低的趋势,分别在24 h、6 h时达到最低值(P0.05),野生蛤仔和白蛤中热休克蛋白家族基因成员表现出先升高后降低的趋势,在胁迫6 h时,白蛤热休克蛋白家族基因成员表达量达到最高值(P0.05),胁迫12 h时,野生蛤仔达到最高值(P0.05);在外套膜组织中,白斑马蛤中热休克蛋白家族基因成员表达量出现先降低后升高的趋势,在胁迫6 h时达到最低值(P0.05),而其他3个壳色蛤仔中热休克蛋白家族基因表达量表现出先升高后降低的趋势,在胁迫6 h时野生蛤仔和白蛤热休克蛋白家族基因成员表达量达到最大值(P0.05),12 h时斑马蛤达到最大值(P0.05),且白蛤HSP40基因表达量在胁迫的各个时间点均显著高于其他3种壳色蛤仔(P0.05)。研究表明,斑马蛤和白蛤的TPx基因可能参与其体内黑色素合成的过程,初步证实了不同壳色蛤仔参与色素合成过程的相关基因在其对低氧的耐受过程中能够发挥重要作用。     

Hydrogen Sulfide Oxidation Is Coupled to Oxidative Phosphorylation in Mitochondria of Solemya reidi

Solemya reidi, a gutless clam found in sulfide-rich habitats, contains within its gills bacterial symbionts thought to oxidize sulfur compounds and provide a reduced carbon food source to the clam. However, the initial step or steps in sulfide oxidation occur in the animal tissue, and mitochondria isolated from both gill and symbiont-free foot tissue of the clam coupled the oxidation of sulfide to oxidative phosphorylation [adenosine triphosphate (ATP) synthesis]. The ability of Solmya reidi to exploit directly the energy in sulfide for ATP synthesis is unprecedented, and suggests that sulfide-habitat animals that lack bacterial symbionts may also use sulfide as an inorganic energy source.     

菲律宾蛤仔糖蛋白提取及体外抗肿瘤活性研究

以菲律宾蛤仔肉为研究对象,水提醇沉,Sevag法除蛋白,DEAE—SepharoseFF阴离子交换层析柱分离纯化,SephadexG-25凝胶柱除盐,得到1种白色组分,红外显示该组分具有糖蛋白的一般吸收特征。将该糖蛋白作用于前列腺癌细胞DU-145进行体外药物敏感性实验,结果显示,0．1moFL氯化钠溶液的洗脱峰组分对DU-145细胞具有明显增殖抑制作用。最适作用浓度约为25mg／mL,表明菲律宾蛤仔具有潜在的抗肿瘤作用。     

背角无齿蚌净化精养池塘水质试验

【目的】探讨背角无齿蚌对池塘养殖水中悬浮颗粒物的滤除效果及其对水体溶解氧（DO）和pH值等的影响,为合理利用该滤食性蚌类构建和优化池塘内部水质净化系统提供科学依据。【方法】在室内的试验条件下,采用不同密度配置的圆背角无齿蚌净化精养鱼池水质,通过测定水中固体悬浮颗粒、DO和pH值,比较其净化效果。【结果】放养圆背角无齿蚌10h后对水中固体悬浮物去除率为47.97%～91.87%,但圆背角无齿蚌的呼吸作用及滤食藻类会引起水体DO和pH值降低。【结论】背角无齿蚌对精养鱼池水中悬浮颗粒物具有较好的滤除效果,但必须要掌握合理的吊养密度,既保证对水中悬浮颗粒物有较高滤除效率,又避免对水体DO和pH造成显著影响。     

Clams As recorders of ocean ridge volcanism and hydrothermal vent field activity

The clam Calyptogena magnifica lives at abyssal depths in association with hydrothermal venting on midocean ridges. Analysis of strontium/calcium ratios in C. magnifica shells provides a temperature proxy with submonthly time resolution. A 21-year strontium/calcium record of two clams from 9 degrees50'N on the East Pacific Rise captures the known 1991 and 1992 eruptive events, documents several additional events between 1992 and 1996, and demonstrates the absence of major hydrothermal episodes during the period 1974 to 1991. These clam archives can increase our understanding of the thermal and chemical history of midocean ridge hydrothermal and volcanic activity on decadal time scales.     

背角无齿蚌净化精养池塘水质试验

【目的】探讨背角无齿蚌对池塘养殖水中悬浮颗粒物的滤除效果及其对水体溶解氧（DO）和pH值等的影响,为合理利用该滤食性蚌类构建和优化池塘内部水质净化系统提供科学依据。【方法】在室内的试验条件下,采用不同密度配置的圆背角无齿蚌净化精养鱼池水质,通过测定水中固体悬浮颗粒、DO和pH值,比较其净化效果。【结果】放养圆背角无齿蚌10 h后对水中固体悬浮物去除率为47.97%～91.87%,但圆背角无齿蚌的呼吸作用及滤食藻类会引起水体DO和pH值降低。【结论】背角无齿蚌对精养鱼池水中悬浮颗粒物具有较好的滤除效果,但必须要掌握合理的吊养密度,既保证对水中悬浮颗粒物有较高滤除效率,又避免对水体DO和pH造成显著影响。     

Oxygen consumption and pumping rates in the hard clam Mercenaria mercenaria: a direct method

Respiratory and pumping rates in the hard clam Mercenaria mercenaria were measured directly in running seawater; the quiatntity of oxygen used was calculated from the difference in concentrations in incurrent and excurrent water. A linear relationship between pumping and respiratory rates suggests regulation of water transport by, oxygen requirement rather than by feeding.     

Hemoglobin in a nonleguminous plant, parasponia: possible genetic origin and function in nitrogen fixation

A dimeric hemoglobin was purified from nitrogen-fixing root nodules formed by association of Rhizobium with a nonleguminous plant, Parasponia. The oxygen dissociation rate constant is probably sufficiently high to allow Parasponia hemoglobin to function in a fashion similar to that of leghemoglobin, by oxygen buffering and transport during symbiotic nitrogen fixation. The identification of hemoglobin in a nonlegume raises important questions about the evolution of plant hemoglobin genes.     

Extended Oxygen Delivery from the Nerve Hemoglobin of Tellina alternata (Bivalvia)

An oxygen-binding hemoglobin localized in the nerves of Tellina alternata (Bivalvia) required 30 minutes to unload oxygen when excised nerves were exposed to pure nitrogen. Neural excitability under these conditions could be sustained only until deoxygenation of the hemoglobin was complete. When the oxygen-combining function of the hemoglobin was abolished with carbon monoxide, the neural excitability ceased within a few minutes of oxygen removal, a response identical to that of hemoglobinless homologous nerves of other bivalves. These results demonstrate that aerobic activity can be supported by the oxygen stored on hemoglobin in microscopic tissues for a considerable time under anoxic conditions.     

Oxygenation properties of snake hemoglobin

Natrix taxispilota hemoglobin has a very high oxygen affinity which depends upon pH in an unusual manner. The oxygen affinity increases slightly upon protein dilution, but the pK's of the Bohr groups are unchanged. Oxidation promotes hemoglobin polymerization, which can be inhibited by prior treatment with iodoacetamide. Reaction with iodoacetamide also causes a slight increase in the oxygen affinity, no change in the pK's of the Bohr groups, and a drastic reduction in heme-heme interaction.     

Erythroid progenitors circulating in the blood of adult individuals produce fetal hemoglobin in culture

Erythroid colonies, raised from erythroid stem cells circulating in the peripheral blood of normal adult individuals, synthesize considerable amounts of fetal hemoglobin. In cultures from persons with sickling disorders, amounts of hemoglobin F that are known to inhibit sickling in vivo are produced. The results provide evidence that primitive erythroid progenitors are able to express the hemoglobin F production program and that cultures of mononuclear cells of the adult blood can be used to investigate the mechanisms involved in regulation of gamma-globin gene switching.     

Hemoglobin switching in sheep and goats: occurrence of hemoglobins A and C in the same red cell

Sheep and goats switch from the synthesis of hemoglobin A (alpha(2)beta(2)(A)) to hemoglobin C (alpha(2)beta(2)(C)) when made anemic. We have demonstrated the existence of the asymmetrical hybrid hemoglobin, alpha(2)beta(A)beta(C), in the circulating red cells of anemic sheep. These erythroid cells, therefore, synthesized both A and C hemoglobin simultaneously. Thus, the switch appears to be mediated by selective gene expression rather than by a clonal or cellular selective mechanism.     

Transport of gases through hemoglobin solution

A mathematical theory is developed to explain the observed enhancement of oxygen transport through solutions by hemoglobin. At high partial pressures of oxygen, ordinary diffusion through the solvent accounts for all transport of oxygen, but at low partial pressures the transport may be increased many fold by the presence of hemoglobin. This phenomenon is explained and the possible role of this phenomenon in living organisms is discussed. The theory also indicates a new method of determining dissociation curves from diffusion experiments.     

缢蛏C型凝集素基因ScCTL-2的序列分析及凝菌活性

为研究缢蛏C型凝集素基因ScCTL-2的结构和功能特点,克隆了该基因的全长序列,并对其mRNA的表达模式以及重组蛋白的凝菌活性进行研究。结果显示,ScCTL-2的cDNA全长2 194 bp,共编码630个氨基酸,其氨基酸序列拥有4个C型凝集素糖识别结构域和1个N端跨膜区,这一特殊的序列结构不同于任何一种已知的C型凝集素。基因mRNA在健康缢蛏肝胰腺中的表达量显著高于其他组织,在鳃中的表达量次之,而在其余组织中的表达量没有显著性差异;金黄色葡萄球菌与鳗弧菌感染能够显著上调ScCTL-2在缢蛏血细胞中的表达。ScCTL-2的重组蛋白凝菌实验显示:在Ca~(2+)存在的情况下,rScCTL-2能够对实验所采用的金黄色葡萄球菌、枯草芽孢杆菌、大肠杆菌、藤黄微球菌和酿酒酵母均产生明显的凝集反应;当溶液中不存在Ca~(2+)时,凝集反应几乎不能发生,意味着rScCTL-2的凝菌活性是严格Ca~(2+)依赖的。研究表明,ScCTL-2可能是一种新的无脊椎动物C型凝集素类型,它具有无脊椎动物C型凝集素普遍具备的免疫活性,可能在缢蛏的免疫中具有重要作用。     

Inheritance of Diego blood group in Mexican Indians

Electron exchange involving the valence states of iron occurs via water bridges. Molecular oxygen reversibly displaces the sole coordinated water of the ferrous iron in hemoglobin, and, in the absence of this ready path for electron transfer, the oxygen is transported without oxidation of the ferrous iron of hemoglobin.     

木瓜蛋白酶酶解制备红岛蛤蜊肉多肽工艺的优化

为获得具有海鲜风味的小分子肽用于功能性食品或调味料中,以红岛蛤蜊肉为原料,综合氨基态氮含量和腥味等指标,筛选出适宜的酶解工艺,并制备出风味鲜美,营养卫生的酶解液。结果表明:经成分测定,蛤蜊肉中脂肪含量为20.01g/100g,蛋白质为30.36g/100g,属于高蛋白低脂肪水产品;利用木瓜蛋白酶对红岛蛤蜊肉进行酶解的最佳工艺为酶解时间4h、木瓜蛋白酶加酶量8 000U/g、酶解温度65℃、初始pH 7.0,该条件下红岛蛤蜊肉酶解液中氨基态氮含量为0.430g/100mL,得到的酶解液颜色淡黄、腥味较小、酶解液澄清、无细小颗粒物质,可作为风味调料的原料应用。     

大竹蛏高通量转录组测序数据组装和分析

为进一步开发大竹蛏Solen grandis的基因资源,采用2代Illumina Hi-seq测序技术对大竹蛏的鳃组织进行了转录组测序,构建了转录组数据库,获得338 483 476条Clean Reads数据;拼接组装后获得190 856条Unigene数据,平均长度为1147 bp;与NR、NT、KO、Swiss Prot、PFAM、GO、KOG等数据库进行Blast信息比对(E-value为10-5),共获得63 337个注释基因;与NR数据库比对发现,大竹蛏转录组基因序列与长牡蛎Crassostrea gigas具有较高的同源性,为53.3%;将大竹蛏转录组的Unigene的功能通过与KOG数据库进行注释比对划分为25类;GO数据库注释可分为三类,即细胞组分、生物过程和分子功能,共包括65个分支;KEGG分析发现,大竹蛏转录组数据中按照代谢通路可分为92类,利用Blast蛋白库比对和Estscan软件进行ORF预测,获得长度大于300 nt的ORF共50 681个;通过SSR分析,共获得73 089个SSR标记。本研究中获得的转录组信息可为今后进行大竹蛏分子标记的开发和关键基因的克隆及功能分析等研究提供基础数据。