The Common Bean Small Heat Shock Protein Nodulin 22 from Phaseolus vulgaris L. Assembles into Functional High-Molecular-Weight Oligomers

Small heat shock proteins (sHsps) are present in all domains of life. These proteins are responsible for binding unfolded proteins to prevent their aggregation. sHsps form dynamic oligomers of different sizes and constitute transient reservoirs for folding competent proteins that are subsequently refolded by ATP-dependent chaperone systems. In plants, the sHsp family is rather diverse and has been associated with the ability of plants to survive diverse environmental stresses. Nodulin 22 (PvNod22) is an sHsp of the common bean (Phaseolus vulgaris L.) located in the endoplasmic reticulum. This protein is expressed in response to stress (heat or oxidative) or in plant roots during mycorrhizal and rhizobial symbiosis. In this work, we study its oligomeric state using a combination of in silico and experimental approaches. We found that recombinant PvNod22 was able to protect a target protein from heat unfolding in vitro. We also demonstrated that PvNod22 assembles into high-molecular-weight oligomers with diameters of ~15 nm under stress-free conditions. These oligomers can cluster together to form high-weight polydisperse agglomerates with temperature-dependent interactions; in contrast, the oligomers are stable regarding temperature.     

Functional Characterization of an Arabidopsis Profilin Protein as a Molecular Chaperone under Heat Shock Stress

Profilins (PFNs) are actin monomer-binding proteins that function as antimicrobial agents in plant phloem sap. Although the roles of Arabidopsis thaliana profilin protein isoforms (AtPFNs) in regulating actin polymerization have already been described, their biochemical and molecular functions remain to be elucidated. Interestingly, a previous study indicated that AtPFN2 with high molecular weight (HMW) complexes showed lower antifungal activity than AtPFN1 with low molecular weight (LMW). These were bacterially expressed and purified to characterize the unknown functions of AtPFNs with different structures. In this study, we found that AtPFN1 and AtPFN2 proteins have LMW and HMW structures, respectively, but only AtPFN2 has a potential function as a molecular chaperone, which has never been reported elsewhere. AtPFN2 has better protein stability than AtPFN1 due to its higher molecular weight under heat shock conditions. The function of AtPFN2 as a holdase chaperone predominated in the HMW complexes, whereas the chaperone function of AtPFN1 was not observed in the LMW forms. These results suggest that AtPFN2 plays a critical role in plant tolerance by increasing hydrophobicity due to external heat stress.     

Progress in Molecular Chaperone Regulation of Heat Shock Protein 90 and Cancer

Heat shock protein 90 (HSP90) is a member of genetically conserved heat shock protein family. As an important molecular chaperone in eukaryotic cells, HSP90 plays a key regulatory role in maintaining cellular protein homeostasis. HSP90 clients encompass a wide range of proteins, thus HSP90 is involved in diverse biological process. With the deeper study, it is found that HSP90 takes an important part in the development and metastasis of cancer, and has become a promising target for the study of anticancer biology. In this review, the progress of HSP90 as molecular chaperone and its relationship with cancer are discussed.     

顺磁共振和傅里叶变换红外光谱法研究高温对荒漠苔藓质膜结构影响

运用顺磁共振波谱和原位傅立叶变换显微红外光谱研究了高温对荒漠植物刺叶墙藓不同叶龄水合组织质膜透性和膜蛋白二级结构的影响。自旋标记法研究质膜透性结果表明,处理前野生叶质膜透性普遍高于室内培养获得的原丝体和次生叶,处理后次生叶膜透性变化最大,比处理前增加4倍,其次为原丝体。质膜透性随叶龄增加,变化幅度逐渐降低。红外光谱的二阶求导、傅立叶自解卷积及拟合分峰等结果显示,各叶龄间蛋白质二级结构含量差异较大,表明各龄组织蛋白成分不完全相同;高温处理后次生叶和原丝体α螺旋含量分别比对照增加40%和16%;其它叶龄组织二级结构含量变化范围小,表明热胁迫下老龄组织蛋白质二级结构稳定。膜透性和膜蛋白二级结构分析证明,膜透性和蛋白质稳定性呈正相关,指示蛋白质组成和含量不同是造成各龄组织不同耐热性的主要原因之一。     

Lysosomal pH Rise during Heat Shock Monitored by a Lysosome‐Targeting Near‐Infrared Ratiometric Fluorescent Probe

Heat stroke is a life‐threatening condition, featuring a high body temperature and malfunction of many organ systems. The relationship between heat shock and lysosomes is poorly understood, mainly because of the lack of a suitable research approach. Herein, by incorporating morpholine into a stable hemicyanine skeleton, we develop a new lysosome‐targeting near‐infrared ratiometric pH probe. In combination with fluorescence imaging, we show for the first time that the lysosomal pH value increases but never decreases during heat shock, which might result from lysosomal membrane permeabilization. We also demonstrate that this lysosomal pH rise is irreversible in living cells. Moreover, the probe is easy to synthesize, and shows superior overall analytical performance as compared to the existing commercial ones. This enhanced performance may enable it to be widely used in more lysosomal models of living cells and in further revealing the mechanisms underlying heat‐related pathology.     

Role of Regulatory Peptide in Pathogenesis of Shock

The present study evaluated the pathogenetic roles of three kinds of regulatory peptide. The results showed that (i) plasma endothelin(ET) level elevated significantly in septic shock rats, persistent intravenous drip of low doses ET caused development of shock state in normal rats and the irreversible outcome of light hemorrhagic shock. Furthermore, i. v. administration of specific ET-antiserum was significantly effective to septic shock rats, (Ⅱ) Plasma calcitonin gene-related peptide (CGRP) increased by 260% in septic shock rats, i. v. drip of low doses CGRP both in early and late sepsis were effective to shock rats, (Ⅱi) An-giotensin-Ⅱ (ANG-Ⅱ) contents of heart and aorta increased dramatically both in early and late septic shock, and inhibiting its increase with Captopril in late sepsis significantly improved the shock state, but results were inverse in early sepsis. It could be concluded that ET was one of the most important factors participating in the pathogenesis of shock, CGRP had a compens     

氨基酸与蛋白质体系热容研究

氨基酸与蛋白质都是生命现象和分子生物学研究的最基本和最重要的研究对象.利用热力学方法,特别是从热容的角度出发对其研究,对深入了解蛋白质的折叠与伸展、变性机理、稳定性及生命体的新陈代谢等问题均具有一定的意义.近年来,研究者们对蛋白质或氨基酸体系的热容研究做了大量工作,取得了很大进展,本文对此进行了概述。     

Heat Effects in ZLC Experiments

The problem of nonisothermal desorption in a zero length column (ZLC) experiment is considered theoretically. Simple analytical expressions for the ZLC desorption curve are derived for certain limiting situations in which the governing equations reduce to a linear form. More general numerical solutions are calculated for a wide range of experimental conditions assuming both negligible mass transfer resistance and finite mass transfer resistance controlled by intraparticle diffusion. A simple criterion for negligible thermal effects is developed. It is shown that when the ZLC technique is applied to the measurement of diffusion in unaggregated zeolite crystals, as originally intended, heat effects are generally insignificant. However, when applied to the measurement of macropore diffusion in relatively large adsorbent particles heat effects can become important and may cause major modification of both the desorption rate and the shape of the desorption curve. A recent experimental ZLC study carried out with commercial adsorbent particles, under conditions of macropore diffusion control, showed an anomalous dependence of the desorption rate on both temperature and particle size. These effects can be qualitatively explained by the nonisothermal model. A more precise quantitative representation of these experiments will require a more refined model incorporating a nonlinear equilibrium isotherm as well as intraparticle diffusional resistance.     

Circulating Ligands of the Receptor for Advanced Glycation End Products and the Soluble Form of the Receptor Modulate Cardiovascular Cell Apoptosis in Diabetes

We determined whether plasma concentrations of the receptor for advanced glycation end products (RAGE) and the soluble (s) form of RAGE (sRAGE) in healthy individuals and patients with type 2 diabetes (T2D) modulate vascular remodeling. Healthy individuals and patients with T2D were divided into two age groups: young = <35 years old or middle-aged (36–64 years old) and stratified based on normal glucose tolerance (NGT), impaired (IGT), and T2D. Plasma titers of sRAGE, the RAGE ligands, AGEs, S100B, S100A1, S100A6, and the apoptotic marker Fas ligand Fas(L) were measured by enzyme-linked immunosorbent assay (ELISA). The apoptotic potential of the above RAGE ligands and sRAGE were assessed in cultured adult rat aortic smooth muscle cells (ASMC). In NGT individuals, aging increased the circulating levels of AGEs and S100B and decreased sRAGE, S100A1 and S100A6. Middle-aged patients with T2D presented higher levels of circulating S100B, AGEs and FasL, but lower levels of sRAGE, S100A1 and S100A6 than individuals with NGT or IGT. Treatment of ASMC with either AGEs or S100B at concentrations detected in T2D patients increased markers of inflammation and apoptosis. Responses attenuated by concomitant administration of sRAGE. In middle-aged patients with T2D, lower circulating plasma levels of sRAGE may limit decoy and exogenous trapping of deleterious pro-apoptotic/pro-inflammatory RAGE ligands AGEs and S100B, increasing the risk for diabetic complications.     

Impaired Chaperone Activity of Human Heat Shock Protein Hsp27 Site‐Specifically Modified with Argpyrimidine

Non‐enzymatic posttranslational modifications (nPTMs) affect at least ～30 % of human proteins, but our understanding of their impact on protein structure and function is limited. Studies of nPTMs are difficult because many modifications are not included in common chemical libraries or protein expression systems and should be introduced site‐specifically. Herein, we probed the effect of the nPTM argpyrimidine on the structure and function of human protein Hsp27, which acquires argpyrimidine at residue 188 in vivo. We developed a synthetic approach to an argpyrimidine building block, which we then incorporated at position 188 of Hsp27 through protein semisynthesis. This modification did not affect the protein secondary structure, but perturbed the oligomeric assembly and impaired chaperone activity. Our work demonstrates that protein function can be altered by a single nPTM and opens up a new area of investigation only accessible by methods that allow site‐selective protein modification.     

CeO2对等离子喷涂Cr2O3涂层抗热震性的影响

在Ｃｒ２Ｏ３陶瓷粉末中添加不同量的ＣｅＯ２,探讨ＣｅＯ２对等离子喷涂涂层抗热震性的影响,认为涂层的热震失效机制是在循环应力作用下发生在陶瓷层／粘结层界面的疲劳失效。在等离子喷涂Ｃｒ２Ｏ３涂层中添加３％ＣｅＯ２后,热震超裂寿命及失效寿命均得到明显提高。适量的ＣｅＯ２使微型裂纹呈现网状分布于涂层薄片内,具有释放涂层内应力的作用,可延缓裂纹的起裂和扩展,并使涂层内贯穿性孔洞减少,从而显著提高涂层的抗热震     

Heat capacity of solid‐state biopolymers by thermal analysis

Heat capacities in the solid state of four globular proteins (bovine β‐lactoglobulin, chicken lysozyme, ovalbumine, and horse myoglobin) and of the poly(amino acid) poly(L ‐tryptophan) have been determined using the Advanced THermal Analysis System (ATHAS). The experimental measurements were performed with adiabatic and differential scanning calorimetry over wide temperature ranges. The heat capacities were linked to an approximate vibrational spectrum by making use of known group vibrations and of a set of parameters, Θ₁ and Θ₃, of the Tarasov function for the skeletal vibrations. Good agreement was found between experiments and calculations with root mean square errors mostly within ±3%. The experimental data were analyzed also with an empirical addition scheme using the known data for poly(amino acid)s measured earlier. Based on this study, vibrational heat capacities can now be predicted for all proteins with an accuracy comparable to common experiments. © 1999 John Wiley & Sons, Inc. J Polym Sci B: Polym Phys 37: 2093–2102, 1999     

硫杂杯芳烃--一类新型的分子受体化合物

综述了近年来一类新型分子受体化合物——硫杂杯芳烃及其衍生物的合成、受体化学以及应用方面的研究进展。     

Heat Transfer Enhancement on the Adsorber of Adsorption Heat Pump

Three methods for improving the heat transfer of the adsorber have been developed in this paper. First, an electrically conductive polyaniline was applied for enhancing the thermal conductivity of adsorbent bed. A thermally conductive composite of polyaniline and adsorbent was prepared by chemical oxidative in situ polymerization of aniline onto the surface of adsorbent particles. A thin thermal conducting net on the surface of the adsorbent particles was grown. The experimental results indicated that the thermal conductivity of this composite could be increased to approximately 4 times that of the raw adsorbent. Second, the adsorbent bed was shaped by a compressing process. This process can reduce the thermal resistance among the adsorbent particles and the contact thermal resistance between the adsorbent bed and the heat exchanger. The thermal conductivity of the shaped adsorbent bed itself from the tests can be increased 30% when the density of the solid adsorbent bed is 1.5 times that of its original density. Furthermore, the adsorption capacity of the above treated adsorbent did not decrease obviously. Third, a proper design of adsorber has been introduced and analyzed. Further tests of this design will be conducted soon.     

关于Mayer关系式和热容比关系式的再讨论

对热力学中关于均匀物质的等压热容(Cp)和等容热容(CV)的2个重要关系式(Mayer关系式和热容比关系式)进行了比较全面的分析和讨论,总结了在教学过程中应该注意的相关事项,以期对教和学都有所裨益。     

Spotlight on Nociceptin/Orphanin FQ Receptor in the Treatment of Pain

In our society today, pain has become a main source of strain on most individuals. It is crucial to develop novel treatments against pain while focusing on decreasing their adverse effects. Throughout the extent of development for new pain therapies, the nociceptin/orphanin FQ receptor (NOP receptor) has appeared to be an encouraging focal point. Concentrating on NOP receptor to treat chronic pain with limited range of unwanted effects serves as a suitable alternative to prototypical opioid morphine that could potentially lead to life-threatening effects caused by respiratory depression in overdose, as well as generate abuse and addiction. In addition to these harmful effects, the uprising opioid epidemic is responsible for becoming one of the most disastrous public health issues in the US. In this article, the contributing molecular and cellular structure in controlling the cellular trafficking of NOP receptor and studies that support the role of NOP receptor and its ligands in pain management are reviewed.     

The Effects of Various Heat Treating Parameters on the Hardness and Microstructures of the Experimental 18% Nickel Maraging Steels

The effect of the parameters of heat treatment on the experimental 18% maraging steels was studied using hardness tester, optical, scanning electron and transmission electron microscopy and X-ray metallography techniques. The specimens were solution treated at 815, 900, 1000, 1060°C for periods between 1 and 4 h. After air-cooling to room temperature, a bcc martensitic structure was obtained. The higher temperature (1060°C) and longer time (4 h) of the solution treatment caused essentially homogeneous, massive martensitic structure. The aging of the steels was studied between 240 and 480°C from 1 to 41 h. As indicated by the results, the hardness vs. aging time curves show a rapid rate of hardening at 480°C while the response at 240 to 320°C is slower. The time required to reach peak hardness increases with decreasing temperature. The increase in hardness during aging can be explained by the precipitation of hardening phases. So, it is necessary to use a high temperature solution treatment to obtain a better alloy distribution and a tough martensitic structure, and an aging treatment at 480 °C between 4 and 10 h to achieve the desired properties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Development of a One-Shot Optical Citrate Sensor Based on a Guanidinium Synthetic Receptor

A new optical absorption-based one-shot sensor for the determination of citrate is described. The citrate-sensitive element is a plasticized polymeric membrane containing a synthetic guanidinium receptor and a pH-sensitive lipophilic dye. The rigid amide-based receptor with C₃ symmetry controls the anion selectivity pattern of the optical element. The optical selectivity coefficients obtained for citrate over a variety of common carboxylate anions meet the requirements for the determination of citrate in different types of samples. The one-shot sensor responds rapidly – in 40 s – and reversibly to citrate over a dynamic range of 0.779 to 25.0 mM with a sensor-to-sensor reproducibility of 5.7%, as log a_cit, in the medium level of the range. The performance of the optical one-shot sensor has been tested in the analysis of citrate in soft drinks and pharmaceuticals, validating the results against a reference procedure. The proposed method is quick, inexpensive, selective and sensitive, and uses conventional instrumentation only.     

膜受体介导双酚A低剂量内分泌干扰效应的分子机制

双酚A(Bisphenol A, BPA)为苯酚系衍生物,作为生产聚碳酸酯塑料和环氧树脂的中间体被广泛应用于多种产品制造中,为全球产量最大的工业品之一。BPA的大量应用使普通人群暴露BPA的几率显著增加。BPA结构与雌激素相似,为一典型雌激素样内分泌干扰物,可以对机体产生多种毒性效应。高剂量BPA主要通过拮抗雌激素受体而发挥其内分泌干扰效应;环境相关低剂量BPA由于不能与雌激素竞争结合雌激素受体,主要通过膜受体介导的信号通路以非基因组方式诱导细胞生物学功能改变。但是,具体何种膜受体介导BPA的低剂量效应以及相关分子机制目前还不清楚。基于此,我们课题组近年来在这些方面做了一系列工作。我们发现,膜G蛋白偶联受体30和整合素αvβ3及其介导的信号传导通路分别介导了环境相关低剂量BPA对雄性生殖细胞的增殖诱导和甲状腺素基因转录的干扰。对环境相关低剂量BPA作用机制的深入理解不仅有助于更客观真实评价和预测环境暴露BPA对人体健康的可能潜在影响以及采取有针对性的预防和干预措施。同时,也将为评价其他类似结构雌激素样环境内分泌干扰物的健康效应提供理论基础及技术支持。本文将结合我们近年来的研究工作,综述目前环境低剂量BPA暴露对人体健康影响的分子机制研究进展、存在的问题以及将来研究的一些思考。