热激蛋白70的研究进展

热激蛋白(HSP70)作为分子伴侣参与细胞内许多重要反应,从而对生物体起着重要的作用。随着研究的深入,其生物学功能不断被发现和利用的同时,HSP70的应用前景也变得越来越广泛。已有研究者对HSP70的生物学功能做了详细介绍,我们主要对近年来HSP70在医学及环境监测等方面的应用进行综述。     

热休克蛋白对细胞凋亡的调控作用

热休克蛋白属于细胞内分子伴侣蛋白,除涉及细胞内一些蛋白质分子构象和稳定性的调节之外,热休克蛋白对细胞应激、代谢、增殖以及凋亡等生理过程均具有重要的调控作用。研究表明热休克蛋白对细胞凋亡的调控机制是复杂的,可直接作用于与凋亡相关的蛋白质,也可以通过影响细胞信号传递而间接影响凋亡的发生。由于热休克蛋白对细胞凋亡的调控机制大多依赖于其分子伴侣功能,阻断热休克蛋白的伴侣功能已经成为研究药物诱导肿瘤细胞凋亡的重要靶点。     

玉米胚发育过程中热激蛋白的合成

35S-Met标记玉米胚蛋白合成结果表明,热激处理(42℃)与对照(25℃)的蛋白合成趋势相近,热激抑制16 DAP的蛋白合成,增加22和34 DAP蛋白合成.SDS-PAGE自显影图谱表明,热激诱导16DAP的胚合成86.4、80.0、73.2 kD等3种分子量较高的热激蛋白,22DAP后热激诱导合成86.4、80.0、73.2、24.4、18.2、16.8和13.6 kD等7种分子量的热激蛋白.2D-PAGE自显影图谱进一步显示,热激诱导22和28 DAP的胚合成近20种热激蛋白,其中超过10种为小分子热激蛋白.特异热激蛋白BiP(HsP70)、PDI(HsP60)Western blot表明,这2种热激蛋白在玉米胚发育过程均有高水平的表达,热激对其合成影响不明显.     

The Diverse Functions of Small Heat Shock Proteins in the Proteostasis Network

The protein quality control (PQC) system maintains protein homeostasis by counteracting the accumulation of misfolded protein conformers. Substrate degradation and refolding activities executed by ATP-dependent proteases and chaperones constitute major strategies of the proteostasis network. Small heat shock proteins represent ATP-independent chaperones that bind to misfolded proteins, preventing their uncontrolled aggregation. sHsps share the conserved α-crystallin domain (ACD) and gain functional specificity through variable and largely disordered N- and C-terminal extensions (NTE, CTE). They form large, polydisperse oligomers through multiple, weak interactions between NTE/CTEs and ACD dimers. Sequence variations of sHsps and the large variability of sHsp oligomers enable sHsps to fulfill diverse tasks in the PQC network. sHsp oligomers represent inactive yet dynamic resting states that are rapidly deoligomerized and activated upon stress conditions, releasing substrate binding sites in NTEs and ACDs Bound substrates are usually isolated in large sHsp/substrate complexes. This sequestration activity of sHsps represents a third strategy of the proteostasis network. Substrate sequestration reduces the burden for other PQC components during immediate and persistent stress conditions. Sequestered substrates can be released and directed towards refolding pathways by ATP-dependent Hsp70/Hsp100 chaperones or sorted for degradation by autophagic pathways. sHsps can also maintain the dynamic state of phase-separated stress granules (SGs), which store mRNA and translation factors, by reducing the accumulation of misfolded proteins inside SGs and preventing unfolding of SG components. This ensures SG disassembly and regain of translational capacity during recovery periods.     

小热休克蛋白的结构和功能

小热休克蛋白（small heat shock protein,sHSP）几乎存在于所有生物体中,其主要结构是一个保守的α晶体蛋白(α-crystallin)结构域,由约90个氨基酸残基组成,与其相邻的是可变的N端域. N端域能够调节低聚体形成、亚单位动力学及其与底物的结合.sHSP能够与细胞内各组分（蛋白质、细胞核、细胞骨架元件、膜）进行相互作用,以维持细胞的稳定.小热休克蛋白家族成员的共同特点是特殊丝氨酸残基上的磷酸化,磷酸化作用对于受到胁迫时的细胞非常重要.由MAPKAP激酶 2/3和p38参与的级联反应能够诱导sHSP发生磷酸化,从而调节sHSP的低聚体状态,而低聚体状态和sHSP的生物学功能密切相关.本文介绍sHSP的结构特征和细胞内的作用底物,并讨论sHSP被不同的蛋白激酶磷酸化及磷酸化作用对低聚体状态和伴侣活性的影响.     

Thermotolerance in Fungi: The Role of Heat Shock Proteins and Trehalose

The parallel synthesis of heat shock proteins and trehalose in response to heat shock did not allow the role of these compounds in the acquisition of thermotolerance by fungal cells to be established for a long time. This review analyses experimental data obtained with the use of mutant fungal strains and shows differences in the thermoprotective functions of trehalose and heat shock proteins in relation to cell membranes and macromolecules. The main emphasis has been placed on data demonstrating the thermoprotective role of trehalose in fungi, the present-day understanding of its biological functions, and mechanisms of trehalose interaction with subcellular structures and cell macromolecules.__________Translated from Mikrobiologiya, Vol. 74, No. 3, 2005, pp. 293–304. p ]Original Russian Text Copyright © 2005 by Tereshina.     

极端嗜热古菌的热休克蛋白

随着生物工程产业对于耐高温酶和菌体的需求, 极端嗜热古菌热休克蛋白(heat shock proteins, HSPs)的研究更受重视, 其热休克蛋白体系非常简洁, 不含HSP100s和HSP90s, 就是HSP70(DnaK)、HSP40、(DnaJ)和GrpE等嗜温古菌可能含有的在极端嗜热古菌中几乎不含有, 即仅包括HSP60, sHSP, prefoldin和AAA+蛋白四大类, 因此对其结构、功能和作用机制的研究在理论和实践上都特别有意义。系统地介绍了这四大类组分的结构、功能和作用机制和协同作用的研究进展, 论述了极端嗜热古菌热休克蛋白的系列研究难点和困惑, 展望了进一步的研究方向和重点。     

Heat Shock Proteins: Functions and Role in Adaptation to Hyperthermia

The results are generalized of many-year studies into the adaptive role of heat shock proteins in different animals, including the representatives of cold- and warm-blooded species that inhabit regions with different thermal conditions. Adaptive evolution of the response to hyperthermia can lead to different results depending on the species. The thermal threshold of induction of the heat shock proteins in desert thermophylic species is, as a rule, higher than in the moderate climate species. In addition, thermoresistant species are often characterized by a certain level of heat shock proteins in cells even at a physiologically normal temperature. Although adaptation to hyperthermia is achieved in most cases without changes in the number of heat shock genes, they can be amplified in some cases in termophylic species. The role of mobile elements in evolution of the heat shock genes was shown and approach was developed for directional introduction of mutations in the promoter regions of these genes.__________Translated from Ontogenez, Vol. 36, No. 4, 2005, pp. 265–273.Original Russian Text Copyright © 2005 by Evgen’ev, Garbuz, Zatsepina.     

热休克蛋白(HSPs)在胚胎发育中作用的研究进展(英文)

在胚胎发育,特别是早期胚胎发育时期,基因转录活跃,蛋白质大量合成,细胞的增殖和分化非常剧烈,细胞的环境在不断变化,细胞对外界刺激十分敏感。这个时期的ＨＳＰｓ变化和作用表现得非常突出。 本文简要总结了近十年有关热休克蛋白在胚胎发育中作用的研究成果。根据胚胎发育对ＨＳＰｓ的依赖性、ＨＳＰｓ基因表达的发育阶段特异性和组织特异性、干扰胚胎发育中ＨＳＰｓ表达程序以后导致胚胎异常发育等现象推测：（１）热休克基因通过参与和／或调控体形形成、肌肉及神经分化,而在胚胎发育中具有看家基因的功能;（２）热休克蛋白作为伴侣分子,通过介导新合成的和／或可逆变性的蛋白质正确折叠、装配、转运及促进不需要的和／或不可逆变性的蛋白质降解,参与胚胎的正常发育和保护胚胎不受不良刺激的影响。这方面的深入研究,必将有助于阐明胚胎发育、细胞增殖、细胞分化和去分化、细胞转化、生物适应性等的分子机理。     

热休克蛋白的生物学作用和转录水平调控研究进展(英文)

热休克蛋白 (HSP)具有广泛的生物学功能 ,其表达方式有两种 :一种是诱导性表达 ,即当生物、细胞受到刺激时才进行表达 ;另一种是组成性表达 ,即在生物、细胞的正常生活、代谢过程中表达。HSP的这两种表达方式意味着HSP基因表达的调控方式和机理不同。本文简要介绍了热休克因子(HSF)的种类、结构及调控HSP基因表达的机理。HSF通过以下 4个步骤调节HSP基因表达 :( 1 )HSF由单体形式变成磷酸化的三聚体形式被激活 ;( 2 )三聚体形式的HSF与HSP基因的热休克元件(HSE)上相邻排列的 3个 5′ GAA 3′结合 ;( 3)与HSF结合后 ,HSE的活化域暴露 ,HSP基因转录 ;( 4 )HSP的mRNA 5′端前导区的特异结构适合于核糖体快速结合和高效翻译。不同生物体内的HSF作用有一定差异 ,功能较为明确的有 :( 1 )对应激信号敏感的HSF1 ;( 2 )对应激信号不敏感 ,对生长、发育、分化信号敏感的HSF2 ;( 3)起抑制HSP基因转录作用的HSF4。还有一些HSF(如HSF3)的作用机制较复杂 ,有待深入研究。此外 ,本文也简要介绍了HSP在衰老、免疫应答、细胞生存和凋亡平衡等中的作用 ,对了解和认识生物生长、发育、衰老、保护、免疫应答及细胞生存和凋亡平衡的分子机制有一定帮助。     

热激蛋白70研究进展

热激蛋白70（HSP70）是广泛存在且高度保守的蛋白,作为伴侣分子能够促进蛋白折叠;HSP70可以通过阻止细胞色素c从线粒体释放,与凋亡诱导因子结合使其不能入核,或者抑制JNK激酶活性调节细胞凋亡;HSP70可以调节细胞周期进程,促进细胞生长,阻止细胞衰老;免疫功能研究表明HSP70是有效的免疫佐剂,可激发抗原特异性的CTL反应,同时细胞外HSP70和膜结合HSP70可激发非特异性免疫反应。     

热激对水稻幼苗耐冷性及热激蛋白合成的诱导

萌发的水稻种子经42℃热激处理后其幼苗的耐冷性明显增强,膜伤害程度降低,脯氨酸含量增加,超氧物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性和抗氧化物质抗坏血酸含量增加,而膜脂过氧化的关键酶脂氧合酶(LOX)活性及其产物丙二醛(MDA)含量下降.并且热激诱导萌发的水稻胚合成78、70、64、60、46、38、24、17、16kD的热激蛋白(HSP),其中属于HSP70的内质网结合蛋白(BiP)的合成与水稻幼苗耐寒性的提高有关.     

钙-钙调素与小麦苗中热激蛋白的诱导

在 34℃热激条件下 ,种子经钙预处理的小麦苗中的钙调素含量随着热激时间的延长而增加 ,热激 90min时达最大值 ,而种子用钙离子螯合剂EGTA预处理的小麦苗中钙调素含量无明显增加。种子用EGTA及钙调素拮抗剂CPZ和TFP预处理的小麦幼苗在 34℃热激时 ,热激蛋白的合成量减少。 4d的小麦幼苗在34℃或 37℃热激条件下 ,能诱导耐热性的获得 ,分别用EGTA、钙离子通道阻断剂易博定、钙调素拮抗剂TFP或CPZ预处理种子后 ,所得幼苗热诱导的耐热性的提高程度有所下降     

Heat Shock Protein Produced by Helicobacter pylori

The cells of Helicobacter pylori were suspended in the medium containing³⁵S-methionine. After a heat shock of the cells at 42 C for 5, 10, and 30 min, the production of proteins was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Out of many proteins produced by the cells, only 66 kDa protein production was dramatically increased by heat treatment. The N-terminal amino acid sequence of 66 kDa protein was quite similar to that of 62 kDa and 54 kDa proteins previously suggested as heat shock protein (HSP) of H. pylori based on the reaction with polyclonal and monoclonal antibodies against HSP 60 family proteins produced by other bacteria. Therefore, it was concluded that H. pylori produces the 66 kDa protein as its major heat shock protein which belongs to HSP 60 family.     

人工老化处理的卷心菜种子的热激蛋白合成

高活力卷心菜种子蛋白质合成速率比中等活力和低活力种子高很多。热激处理(42℃)下,蛋白质合成显著下降,但高活力种子的蛋白质合成能力仍然显著高于中等和低活力种子。高活力和中等活力种子主要合成分子量为70 kD和一些小分子量的热激蛋白。在低活力种子中检测不到热激蛋白的合成。4种热激蛋白（1种HSP90和3种HSP70）的Western blot检测结果表明,只有1种热激蛋白（HSP70）与种子活力有关。     

几种热激蛋白在细胞凋亡信号通路中的调控作用

热激蛋白(heat shock proteins, HSPs)作为进化保守的蛋白家族 之一,普遍存在于各种生物体中,并在生物体内发挥着重要的生理功能.大 量的实验证据表明,热激蛋白与细胞凋亡密切相关,参与细胞凋亡信号通 路的多个环节. 近年来有关该领域的研究已获得了重要的突破与进展.一方 面,热激蛋白主要起着抑制细胞凋亡、促进细胞存活的作用;另一方面, 某些热激蛋白又能够作为凋亡蛋白的分子伴侣,促进细胞凋亡,比如HSP70 能够激活DNase来促使细胞凋亡,线粒体内HSP60能够促进caspase依赖的细 胞凋亡途径.本文在阐明细胞凋亡信号通路的基础上,综述了近年来几种不 同热激蛋白家族（HSP90、 HSP70 、HSP60和小分子HSPs）在细胞凋亡调控 中作用的研究进展,重点阐述了几种主要热激蛋白与细胞凋亡信号通路上 相关因子的相互作用,并绘制了热激蛋白在细胞凋亡信号通路中的调控图 ,为进一步完善细胞凋亡调控网络研究提供一定的参考.     

OPG-HSP65重组蛋白的原核表达及其生物学活性研究

PCR扩增OPG-HSP65基因,构建原核重组表达载体pET-28a－OPG-HSP65,转化大肠杆菌BL21（DE3）,经IPTG诱导表达产生包涵体形式的目的蛋白。对重组蛋白进行Western blot检测表明,重组蛋白能与抗His-Tag单克隆抗体及鼠抗人OPG单克隆抗体特异性结合。对重组蛋白进行尿素洗涤纯化,进而透析、复性。经破骨细胞生长抑制实验和抑炎实验表明,重组蛋白能减少破骨细胞生成及减轻迟发型超敏反应小鼠模型炎症反应。     

叶绿体小分子量热激蛋白介绍

本文对叶绿体小分子量热激蛋白的研究进行了简要的回顾和总结.叶绿体小分子量热激蛋白是热激蛋白超家族的成员,具有3个特殊的保守区域;当植物遇到热胁迫时,叶绿体小分子量热激蛋白能够保护光合系统Ⅱ和类囊体膜;初步分析了叶绿体小分子量热激蛋白与植物的耐热性和耐冷性关系以及其分子伴侣功能.     

Two Bacterial Small Heat Shock Proteins,IbpA and IbpB,Form a Functional Heterodimer

Small heat shock proteins (sHsps) are a conserved class of ATP-independent chaperones which in stress conditions bind to unfolded protein substrates and prevent their irreversible aggregation. Substrates trapped in sHsps-containing aggregates are efficiently refolded into native structures by ATP-dependent Hsp70 and Hsp100 chaperones. Most γ-proteobacteria possess a single sHsp (IbpA), while in a subset of Enterobacterales, as a consequence of ibpA gene duplication event, a two-protein sHsp (IbpA and IbpB) system has evolved. IbpA and IbpB are functionally divergent. Purified IbpA, but not IbpB, stably interacts with aggregated substrates, yet both sHsps are required to be present at the substrate denaturation step for subsequent efficient Hsp70-Hsp100-dependent substrate refolding. IbpA and IbpB interact with each other, influence each other’s expression levels and degradation rates. However, the crucial information on how these two sHsps interact and what is the basic building block required for proper sHsps functioning was missing. Here, based on NMR, mass spectrometry and crosslinking studies, we show that IbpA-IbpB heterodimer is a dominating functional unit of the two sHsp system in Enterobacterales. The principle of heterodimer formation is similar to one described for homodimers of single bacterial sHsps. β-hairpins formed by strands β5 and β7 of IbpA or IbpB crystallin domains associate with the other one's β-sandwich in the heterodimer structure. Relying on crosslinking and molecular dynamics studies, we also propose the orientation of two IbpA-IbpB heterodimers in a higher order tetrameric structure.