Heat-shock protein 90 inCandida albicans

Researches on Candidal heat-shock protein 90 (HSP90) in recent years are summarized.Candida albicans is a commensal pathogen in human and animals. In immunocompromised individuals it behaves as an opportunist pathogen, giving rise to superficial or systemic infections. Systemic candidosis is a common cause of death among immunocompromised and debilitated patients, in which the mortality is as high as 70%. HSP90 is now recognized as an immunodominant antigen inC. albicans and plays a key role in systemic candidosis as a molecular chaperone. The 47-ku peptide is the breakdown product of HSP90. Patients who has recovered from systemic candidosis produce high titre of antibodies to 47-ku antigen, whereas the fatal cases have little antibody or falling titres. The three commonest epitopes of candidal HSP90 have been mapped, epitopes C, B and H. Epitopes C and H are immunogenic. The antibody probes of both epitopes may be developed into a new serological test agents for systemic candidosis due to rather high specificity and sensitivity. The recent results establish HSP90 as an ATP-dependent chaperone that is involved in the folding of cell regulatory proteins and in the refolding of stress-denatured polypeptides. Some researches on fungal HSP90 and the treatment of patients with candidosis are reviewed as well.     

Phytochrome controlled, long-day photoperiod-inducible protein in rice leaves

A new protein in the leaves of NK58S and NK58 (Oryza sativa L. subsp.japonica), which can be induced by 10 d-long-day photoperiod (14 h lightid) and cannot be induced by 10 d-short-day photoperiod (10 h light/d), has been found by two-dimensional gel electrophoresis. The protein, whose molecular weight and isoelectric point are 36 ku and pH 5.2 respectively, is found to be controlled by phytochrome as shown by the experiment of red light induction-far red light reversion.The existence of this protein in both NK58S and NK58 reflects that some of the responses of NK58S and NK58 might be similar in response to long-day photoperiod, a mild stress.     

Activities and properties of calcineurin catalytic domain

Calcineurin (CN) is the only protein phosphatase known to be under the control of calcium (Ca²⁺) and calmodulin (CaM). The enzyme consists of two subunits, the catalytic A subunit of 61 ku (CNA) and a regulatory B subunit of 19 ku (CNB). In this study, we used PCR amplication to construct a truncation consisting of only the CNA catalytic domain. The truncation was induced by IPTG and expressed inE. coli. PNPP was used as a substrate to study the phosphatase activity of the CNA catalytic domain. The findings show that its activity is 20 times greater than CNA in the presence of CNB and CaM. The optimum reaction temperature for the CNA catalytic domain protein is 40°C, and the optimum reaction pH value is 8.0. Mn²⁺ is still an effective activator for the CNA catalytic domain, but its activity is not controlled by Ca²⁺. In the presence of 6 mmol/L Mg²⁺, adding either Ca²⁺ or EGTA did not change the activity of the CNA catalytic domain.     

NF-κB相互作用多肽原核表达载体的构建、表达与纯化

为了获得高纯度的可溶性NF-κB相互作用多肽,首先以酵母双杂交技术筛选获得的NF-κB相互作用多肽的酵母表达质粒pGAD GH/pp10为模板,扩增多肽基因片段,后经BglⅡ,StuⅠ双酶切后连接到pET-42a(+)载体中,构建GST/多肽融合蛋白的原核表达载体,并将GST/多肽融合蛋白的原核表达载体转化入BL-21菌株,用0.4 mmol/L IPTG于30 ℃诱导表达4 h,后经GST亲和纯化GST/多肽融合蛋白,SDS-PAGE电泳鉴定融合蛋白的表达和纯度.实验结果表明,成功地构建了NF-κB相互作用多肽的GST/多肽融合蛋白原核表达载体,进行了GST/多肽融合蛋白的诱导表达,融合蛋白的表达为可溶性表达,最终纯化获得纯度约为80%的可溶性GST/多肽融合蛋白,这将为后继利用GST pull-down,Bio-sensor,EMSA等实验进一步验证NF-κB相互作用多肽的功能提供可靠的材料来源奠定基础.     

硝酸还原酶和可溶性蛋白对东北三省蒙古栎种源生长性状的影响

本文研究了东北三省25个蒙古栎种源的硝酸还原酶和可溶性蛋白与生长性状的关系．不同生长性状的蒙古栎种源,硝酸还原酶活性和可溶性蛋白含量上都存在着显著的差异,并且同一种源呈现明显的季节变化．试验结果表明生长性状与所研究的两个理化指标之问存在显著的正相关,即高、径生长好的种源,硝酸还原酶活性和可溶性蛋白含量高;硝酸还原酶活性大的种源,可溶性蛋白含量也高,可以为林木育种提供参考．本试验中,集安种源在两项生理指标中含量均最高,可作为优质种源进行推广．     

一种改进的六联体使用频率编码测度

在基因预测软件中常用的编码测度得到的序列编码潜力大小往往与序列的C G含量紧密相关,从而影响了对蛋白编码区的识别效果．研究发现六联体使用偏好与其自身C G含量存在一种近似线性的相关性,据此提出了一种改进的六联体使用偏好模型,通过综合考虑六联体使用频率与六联体的C G含量,可简便有效地减小序列编码潜力大小对序列C G含量的依赖性．测试表明,与分类建模策略相比,该方法所需的训练数据较少,而且具有更好的蛋白编码区识别效果,因此可用于基因预测软件中以提高蛋白编码区与基因结构的预测精度．     

Aβ_(37-42)形成二聚体的分子模拟研究

阿尔茨海默症是一种神经退行性疾病,多发病于老年人群.该疾病不但严重影响患者的健康及正常生活,还给患者家属带来极大痛苦.因此治疗该疾病成为科学界急欲攻克的难关.而治疗的基础是解析病理,大量研究表明该病患者脑内有大量纤维状Aβ蛋白质聚集物.因此研究Aβ蛋白质如何聚集是解析病理的关键.同时预防和阻止Aβ蛋白质聚集也成为治疗阿尔茨海默症的一种可选方案.本文即选取Aβ蛋白质片段,以分子动力学模拟方法,详细分析其形成稳定二聚体的完整过程.研究发现该蛋白质片段上残基侧链之间的疏水作用促使两个Aβ37-42单体相互靠近,其间会形成多种非稳态中间体,其结构经过不断调整最终才形成稳定的二聚体结构.该结构调整过程对形成稳定的二聚体结构非常重要.在此过程中两条肽链主链形成氢键的个数逐渐增加,同时伴随形成了新的残基侧链之间的相互作用.研究发现在二聚体形成的全过程中Ile41和Ala42均起到不可忽视的作用.Ile41由于其侧链具有疏水性且体积相对较大,可通过其疏水作用力促使两个单体相互靠近.而Ala42在后期结构调整中发挥了一定作用,稳定了二聚体的结构.该研究有助于更好地理解Aβ蛋白质聚集特别是其形成二聚体的过程,同时也为防治Aβ蛋白质聚集提供了一些理论依据.     

Morphometrical analysis of four populations of the Little Blue Penguin,Eudyptula minor

Variations in coloration, external morphology and plasma protein between four populations of the Little Blue Penguin, E. minor were determined. A south-north cline was found along the coast of New Zealand.