红螯螯虾感光器中Gq蛋白的鉴定及光波长对其含量的影响

用蛋白质常规提取方法分别提取红螯螯虾复眼视网膜的可溶性和膜结合蛋白质,用SDS-PAGE和免疫印迹法进行G蛋白的分离和鉴定;在经光、暗适应及红、绿、蓝、黄4种波长光照处理后,对其感光器中G蛋白分别进行SDS-PAGE分析.暗适应、自然光及4种波长光适应的感光器可溶性蛋白、膜结合蛋白条带数量和分子量大小基本相同,在42?kD处都出现一条清晰的蛋白条带.兔抗Gαq-11多克隆抗体能识别这一蛋白条带.表明红螯螯虾感光器中存在可溶性和膜结合Gq蛋白α亚基,分子量为42?kD.在不同光照条件下感光器中可溶性和膜结合Gqα亚基的含量有差异.可溶性Gqα蛋白占总蛋白的百分含量依次为:日光(7.71%)>黄光(7.32%)>红光(7.06%)>暗适应(6.94%)>蓝光(6.46%)>绿光(5.74%);除暗适应与红光之间差异不显著外,红光与黄光差异显著,其余差异极显著.而膜结合Gqα亚基的百分含量则相反,依次为:绿光(13.94%)>蓝光(10.56%)>暗适应(10.25%)>红光(10.14%)>黄光(9.76%)>日光(9.44%);除暗适应与红光差异不显著外,红光与黄光差异显著,其余差异极显著.推测红螯螯虾感光器中Gq蛋白的激活与光照刺激的波长有关,其感光器对不同波长光刺激的敏感度有差别.     

中国前鳃亚纲玉螺科新种记述

中国近海前鳃亚纲玉螺科种类丰富,作者就中国科学院海洋研究所历年采到的标本进行了研究,共整理出6属58种;其中有19种为中国海新纪录,8种为新种。新种的模式标本均保存在中国科学院海洋研究所。 1.海南玉螺(新种)Natica hainanensis sp.nov.(图版Ⅰ∶4;图版Ⅱ∶9) 贝壳中等大小,壳较厚、质坚实,亚球形。壳表有细密的生长线和螺线,有的标本有裂痕。壳表被有一层蜡黄色的外皮。在体螺层上弥散有若干褐色斑点;此斑点在有的标本排列成三条螺带,但较模糊。若除去蜡黄色外皮,贝壳呈灰白色。螺旋部较高而尖,褶皱     

厚壳贻贝低分子量足丝蛋白的初步鉴定(英文)

足丝蛋白是贻贝科(Mytilidae)所特有一种在水环境中也能表现出强黏附功能的蛋白,也是目前开发新型生物黏附剂的主要候选分子。厚壳贻贝(Mytilus coruscus)广泛分布于我国东部沿海,是我国具有重要经济价值的贻贝,其足丝粗硬,黏附力强,关于厚壳贻贝的足丝蛋白的研究目前尚未见报道。通过醋酸抽提结合反相高效液相色谱分离,从厚壳贻贝足丝盘中分离纯化到数种低分子量足丝蛋白,经质谱鉴定和氨基酸序列测定,其中三种足丝蛋白(分子量6 kD左右)属于贻贝足丝蛋白-3(mytilus foot protein-3,mfp-3)家族,且序列中富含DOPA,另有三种足丝蛋白为未知新型足丝蛋白。石英晶体微天平分析表明,厚壳贻贝低分子量足丝蛋白在金表面有较强的吸附能力,这与其黏附功能是直接相关的。以上工作为深入了解厚壳贻贝低分子量足丝蛋白的分子多样性以及黏附机制奠定了基础。     

干旱对4种苗木根尖可溶性蛋白组分和含量的影响

以2年生刺槐、油松、侧柏、沙棘苗木为材料,设置包括适宜水分、中度干旱、重度干旱(分别为田间土壤持水量的70%、50%、35% )3个水分程度和短期(7 d)、中期(14 d)、长期(21 d)3个时间的人为控水胁迫实验,研究了干旱胁迫对4种苗木根尖可溶性蛋白含量及组分的影响.结果表明:刺槐根尖可溶性蛋白含量在短期中度干旱和长期重度干旱下显著减少,而在中长期中度干旱和短期重度干旱下显著增加;侧柏根尖可溶性蛋白含量在短期中度干旱和重度干旱下显著增加,长期中度干旱和中长期重度干旱下减少;随着胁迫程度加重和处理时间的延长,油松根尖可溶性蛋白含量总体增加,而沙棘则减少.4种苗木根尖可溶性蛋白组分在不同干旱胁迫程度和时间下均无明显变化,各分子量可溶性蛋白含量变化大多数与总蛋白含量变化趋于一致.可见,4种苗木根尖可溶性蛋白含量的变化在干旱胁迫程度、胁迫时间以及树种之间差异较大,其对干旱的响应是通过大部分蛋白组分较一致地变化来实现的.     

丽蝇蛹集金小蜂寄生对寄主蛹可溶性蛋白与芳基蛋白组成与含量的影响

为了探讨蛹期寄生蜂对寄主蛋白代谢的寄生生理效应,利用Bradford蛋白含量测定法、Western免疫印迹法及酶联免疫吸附检测法研究了棕尾别麻蝇Boettcherisca peregrina蛹被丽蝇蛹集金小蜂Nasonia vitripennis寄生后其脂肪体和血淋巴中可溶性蛋白及芳基蛋白组成与含量的变化。结果表明：寄生蛹脂肪体和血淋巴中可溶性蛋白的组成与未寄生相比基本无明显差异; 不论寄生与否寄主蛹脂肪体和血淋巴中芳基蛋白亚基分子量均为80 kDa,该亚基在脂肪体中未出现降解现象,而在血淋巴中仅于寄生后12 h的寄主蛹中呈现2条分子量相近的Western免疫印迹带,说明其降解可能先于未寄生对照。就含量而言,寄生蛹脂肪体中可溶性蛋白含量除寄生后24 h外均显著低于未寄生对照,芳基蛋白含量除寄生后48 h外也均显著低于未寄生对照,其中寄生后12 h的含量仅为未寄生的32.0%。寄生蛹血淋巴中可溶性蛋白含量多低于未寄生蛹,且寄生后2,12,24 h的差异达显著水平;芳基蛋白的含量均有低于未寄生的趋势,其中寄生后12 h的含量为未寄生的17.0%。综合认为,丽蝇蛹集金小蜂的寄生可导致寄主脂肪体和血淋巴中可溶性蛋白及芳基蛋白含量下降。     

高温胁迫对Bt棉铃壳中Bt蛋白含量及氮代谢的影响

以Bt基因来源于中国的棉花品种泗抗1 号(常规种)、泗抗3 号(杂交种)和来源于美国的棉花品种99B(常规棉)、岱杂1 号(杂交棉)为材料,研究了不同高温水平下Bt 棉盛铃期铃壳中Bt 蛋白含量变化及氮代谢生理特征.结果表明: 铃壳中Bt 蛋白含量随温度升高而降低,与对照相比(32 ℃),常规棉品种在38 ℃、杂交棉品种在40 ℃以上时,铃壳中Bt 蛋白含量大幅度下降.其中,常规种泗抗1号和99B在38 ℃时分别下降53.0%和69.5%;杂交种泗抗3号和岱杂1号在40 ℃时下降64.8%和54.1%.铃壳Bt 杀虫蛋白含量下降显著时,其可溶性蛋白含量明显下降,游离氨基酸含量明显提高,GPT活性显著下降,蛋白酶活性显著增加.高温影响铃壳的氮代谢引起Bt蛋白的分解加剧,合成减弱,从而造成Bt蛋白含量减少,抗虫性下降.     

我国椎实螺科新纪录

现报道我国椎实螺科4种新纪录,标本均于1965年采自云南省。 1.泰国萝卜螺Radix siamensis(Sowerby) 贝壳呈橄揽形;有4个螺层;体螺层圆胖,有的标本具有方形凹陷。壳口呈卵圆形,内缘在轴缘处外翻,扭转,形成轴折。     

九里香多糖和蛋白多糖的分离、纯化和分析

九里香(Murraya Paniculata L.Jack)皮部经热水提取、乙醇沉淀、SephadexG—100柱层析分离、磷酸氢钙吸附、酸溶,再通过Sephadex G-200柱层析纯化,得到灰白色粉末状九里香蛋白多糖。总糖含量为52.1％(其中葡萄糖醛酸含量为10.9％),蛋白质含量为20.0％。九里香蛋白多糖去蛋白后,即得九里香多糖,总糖含量为88.2％(其中葡萄糖醛酸含量为20.0％),经纸层析和聚丙烯酰胺凝胶园盘电泳鉴定为单一斑点和单一区带。平均分子量约为1.7×10~(-5)。组成单糖的摩尔比为葡萄糖:露甘糖:木糖:阿拉伯糖:岩藻糖:葡萄糖醛酸=1.0:0.40:0.16:0.17:0.20:0.48.     

中国甘肃南部鸟唇螺属(柄眼目,艾纳螺总科)一新种描述

描述了栖息于甘肃南部的陆贝艾纳螺科1新种,齐氏鸟唇螺Petraeomastus qii sp.nov..新种以贝壳壳顶尖出、轴唇倾斜及测量特征与各已知种相区别.齐氏鸟唇螺,新种Petraeomastus qii sp.nov.(图1～3)鉴别特征贝壳呈卵锥状,壳顶尖出,螺轴倾斜.壳高10.8 mm,壳径7.1mm,螺层数6.正模,HBUMM06530-specimen 1,成体;甘肃文县(32°56′N,104°40′E;海拔1 024 m);2011-08-08;吴岷、徐沁、Prem B.Buhda采.标本保存于河北大学博物馆.词源:新种种名源自我国贝类学家齐锺彦教授的姓氏;名词.     

浙江弯螺属一新种记述(肺螺亚纲,柄眼目,扭轴蜗牛科)

记述了浙江弯螺属一新种,即龙塘山弯螺新种Sinoennea longtangshanensis sp.nov.。标本采自浙江省临安市龙塘山清凉峰自然保护区。弯螺属的种类主要分布于东亚和南亚地区,在我国主要分布于长江以南的地区。龙塘山弯螺贝壳小型(壳高4.40 mm,壳宽2.40 mm),壳质薄,有6.5个螺层,胚螺层和第二螺层光滑,无肋纹,壳口具4枚齿。模式标本保存于中国科学院动物研究所。     

The Micro/Nanostructure Characteristics and the Mechanical Properties of Hemifusus tuba Conch Shell

<正> The mollusk shell mobilizes calcium from environment for skeletal mineralization.This occurs through synthesizing solidsin solution in the presence of organic molecules of specific interior regions of the conch shell.The ultrastructure and microhardnessof the Hemifusus tuba conch shell living in the Huang/Bo sea area are investigated in the paper.It is shown that thecomposition and microstructure of the mollusk shell vary in different positions.The prodissoconch shell consists only of aragonitewith the crossed-lamellar microstructure.While the spiral shell and the body shell of the Hemifusus tuba conch shell arecomposed of one calcite layer and several aragonite layers.The calcite layer consists of cylindrical grains,but the aragonitelayers are crossed-lamellar ultrastructure at three size scales.The minimum structure size (the third-order lamella) is at about20 nm - 80 nm.The margin of shell aperture is only composed of calcite with cylindrical grains.This natural optimization of theshell microstructure is intimately due to the growth of the Organic matrix.At different positions the microhardness of molluscshell is different due to different crystal structures and crystal arrangements.The growth process of shells allows a constantrenewal of the material,thus enabling their functional adaptation to external environments.     

Aragonite shells are more ancient than calcite ones in bivalves: new evidence based on omics

Two calcium carbonate crystal polymorphs, aragonite and calcite, are the main inorganic components of mollusk shells. Some fossil evidences suggest that aragonite shell is more ancient than calcite shell for the Bivalvia. But, the molecular biology evidence for the above deduction is absent. In this study, we searched for homologs of bivalve aragonite-related and calcite-related shell proteins in the oyster genome, and found that no homologs of calcite-related shell protein but some homologs of aragonite-related shell proteins in the oyster genome. We explained the results as the new evidence to support that aragonite shells are more ancient than calcite shells in bivalves combined the published biogeological and seawater chemistry data.     

N40, a novel nonacidic matrix protein from pearl oyster nacre, facilitates nucleation of aragonite in vitro

A novel nonacidic matrix protein from pearl oyster nacre has been purified by cation-exchange chromatography. It was designated N40 for the nacreous protein of approximately 40 kDa. On the basis of the extraction method (with Tris-buffered Milli-Q water) and amino acid compositions (Gly- and Ala-rich), N40 was inferred to be a conventional "insoluble matrix protein". Crystallization experiments showed that N40 could facilitate the nucleation of aragonite drastically. So far, among the macromolecules that have been purified from the shell, N40 is an exclusive protein that can nucleate aragonite by itself, without the need for adsorption to a substrate. Thus, the present study has proposed the possibility that the nonacidic shell protein (maybe a conventional "insoluble framework protein") can also directly participate in aragonite nucleation and even act as a nucleation site. It is a valuable supplement to the classic biomineralization theory, in which the soluble acidic proteins of the shell are generally believed to function as a nucleation site.     

A Novel Acidic Matrix Protein,PfN44, Stabilizes Magnesium Calcite to Inhibit the Crystallization of Aragonite

Magnesium is widely used to control calcium carbonate deposition in the shell of pearl oysters. Matrix proteins in the shell are responsible for nucleation and growth of calcium carbonate crystals. However, there is no direct evidence supporting a connection between matrix proteins and magnesium. Here, we identified a novel acidic matrix protein named PfN44 that affected aragonite formation in the shell of the pearl oyster Pinctada fucata. Using immunogold labeling assays, we found PfN44 in both the nacreous and prismatic layers. In shell repair, PfN44 was repressed, whereas other matrix proteins were up-regulated. Disturbing the function of PfN44 by RNAi led to the deposition of porous nacreous tablets with overgrowth of crystals in the nacreous layer. By in vitro circular dichroism spectra and fluorescence quenching, we found that PfN44 bound to both calcium and magnesium with a stronger affinity for magnesium. During in vitro calcium carbonate crystallization and calcification of amorphous calcium carbonate, PfN44 regulated the magnesium content of crystalline carbonate polymorphs and stabilized magnesium calcite to inhibit aragonite deposition. Taken together, our results suggested that by stabilizing magnesium calcite to inhibit aragonite deposition, PfN44 participated in P. fucata shell formation. These observations extend our understanding of the connections between matrix proteins and magnesium.     

Characterization of a novel shell matrix protein with vWA domain from Mytilus coruscus

ABSTRACT

Mollusk shell is a product of biomineralization with excellent mechanical properties, and the shell matrix proteins (SMPs) have important functions in shell formation. A vWA domain-containing protein (VDCP) was identified from the shell of Mytilus coruscus as a novel shell matrix protein. The VDCP gene is expressed at a high level in specific locations in the mantle and adductor muscle. Recombinant VDCP (rVDCP) showed abilities to alter the morphology of both calcite and aragonite, induce the polymorph change of calcite, bind calcite, and decrease the crystallization rate of calcite. In addition, immunohistochemistry analyses revealed the specific location of VDCP in the mantle, the adductor muscle, and the myostracum layer of the shell. Furthermore, a pull-down analysis revealed eight protein interaction partners of VDCP in shell matrices and provided a possible protein–protein interaction network of VDCP in the shell.     

Effect of sex and size on the protein composition in the tissues of the freshwater crab,Paratelphusa (Barytelphusa) guerini Milne Edwards

Soluble (enzymic) and insoluble (structural) proteins were estimated quantitatively in different tissues of the crab as a function of sex and size. The different protein fractions in the tissues of females decreased with weight. But the proteins in the tissues of males increased gradually with size up to 30–40 g and decreased later. In both sexes, the relative increase and decrease varied in different tissues.The insoluble protein content of the muscle was always greater than the soluble content in both sexes. However, the two fractions in other tissues showed different trends depending upon the sex. The soluble content was always greater than the insoluble content in gill, heart and hepatopancreas of males. The hepatopancreas of females had a higher soluble content at all weights, as in males. But the insoluble proteins of the gill and soluble proteins of the heart were more in smaller animals (below 25–30 g) and less in larger animals (above 25–30 g).The soluble proteins dominated in the males. This relation was maintained in the muscle throughout the weight range studed (10 to 75 g), while in other tissues this relation was seen for the the greater part of this weight range. The insoluble proteins in the different tissues tended to be more in smaller females (up to 25–30 g) and larger males (above 25–30 g).The total protein content of the muscle was always larger in males. The proteins of the gill and heart were larger in males at higher sizes (above 20–25 g). Hepatopancreatic proteins were larger only at the intermediate-sized males (between 25 and 60 g).     

正常与患白内障大鼠晶状体中脲溶性蛋白质性质的研究

本文研究了正常及三种类型白内障大鼠晶状体中脲溶性蛋白质的含量及性质的变化,发现在每种类型白内障晶状体中,水溶性蛋白质均减少,水不溶性蛋白质则都相对增加。经SephadexG-200柱层析及SDS聚丙烯酰胺凝胶电泳发现,晶状体中脲溶性蛋白质主要是由二硫键交联而成的高分子聚合物。经巯基乙醇还原后,绝大部分高分子聚合物可分解成低分子量蛋白质,其分子量与水溶性的γ晶体蛋白相同。这提示晶状体中脲溶性蛋白质的主要成分很可能是以二硫键交联而成的γ晶体蛋白聚合物。此结果与本实验室所得白内障晶状体水溶性蛋白质的变化相吻合。     

厚壳贻贝whirlin类贝壳基质蛋白的重组表达与功能分析

贝壳历来是生物工程和材料学研究的重要对象。贝壳中的贝壳基质蛋白质在贝壳的形成与发育过程中具有重要的调控作用。Whirlin类蛋白质(Whirlin-like protein,WLP)是一种从厚壳贻贝(Mytilus coruscus)中鉴定的新型贝壳基质蛋白质。序列分析结果显示,该蛋白质含有PDZ(postsynaptic density/Discs large/Zonula occludens)结构域,而该结构域对贝壳生物矿化的影响目前尚无报道。为深入了解WLP在贝壳形成中对碳酸钙晶体的影响,在序列分析基础上,采用密码子优化结合原核重组表达,获得其重组表达产物后,开展了重组WLP对碳酸钙晶体形貌及晶型的影响研究,结晶速度抑制以及碳酸钙晶体结合分析。分析结果表明,重组WLP能诱导文石型碳酸钙晶体的形貌和方解石型碳酸钙晶体的晶型发生改变;同时重组WLP对碳酸钙晶体具有结合作用,且能抑制碳酸钙晶体的结晶速度。上述结果表明,WLP对贝壳的形成及发育具有重要影响,并可能在贝壳肌棱柱层的形成中发挥了重要作用。     

Direct observation of the transition from calcite to aragonite growth as induced by abalone shell proteins

The mixture of EDTA-soluble proteins found in abalone nacre are known to cause the nucleation and growth of aragonite on calcite seed crystals in supersaturated solutions of calcium carbonate. Past atomic force microscope studies of the interaction of these proteins with calcite crystals did not observe this transition because no information about the crystal polymorph on the surface was obtained. Here we have used the atomic force microscope to directly observe changes in the atomic lattice on a calcite seed crystal after the introduction of abalone shell proteins. The observed changes are consistent with a transition to (001) aragonite growth on a (1014) calcite surface.