蛋白质构象病提示的疾病防治新思路

蛋白质构象病是由于组织中特定的蛋白质承受了构象变化,进而聚集并产生沉积所引起的一种疾病。构象病概念的提出提示人们可以通过抑制或者逆转组织蛋白的变构来防治疾病,本文就蛋白质构象病的概念以及近年关注较多的β折叠形成阻断肽和分子伴侣两种防治思路予以综述。     

蛋白质构象病

蛋白质的空间结构又称为三维结构或构象（conformation）,特定的空间构象是蛋白质发挥其各种功能的结构基础。由于蛋白质担负着复杂的生化反应,因此在生物合成以后,蛋白质本身也经历着复杂的生理过程;蛋白质自翻译以后,需进行一系列的翻译后过程,包括跨膜转运、修饰加工、折叠复性、生化反应、生物降解等,这些过程都伴随着蛋白质的结构转换。随着对疯牛病的研究,人们发现：蛋白质分子的氨基酸序列虽不改变,但其空间结构或构象的改变也能引起疾病。同时,越来越多的研究表明,一些遗传性疾病是由于基因突变导致了蛋白质的错误折叠,这些突变并不直接影响蛋白质的功能结构域,但由于蛋白质的错误折叠,干扰了其正确运输,形成对细胞有毒性作用的聚积物。     

伴侣素的发现者——霍维茨

伴侣素（chaperonin）是辅助蛋白质正确折叠过程中的重要元件,可有效防止蛋白质错误折叠和聚集,对细胞正常功能发挥和发育具有十分重要的意义。伴侣素功能缺陷或异常可导致许多神经退行性疾病如帕金森综合症等的发生,因此伴侣素介导的蛋白质折叠的研究对该类疾病的治疗具有重要帮助。伴侣素于1989年由霍维茨发现,经过近20年研究,人们对其作用机理和生理功能有了较为全面的理解,从而为应用奠定了基础。     

植物内质网胁迫研究进展

内质网是蛋白质折叠和蛋白质糖基化修饰的重要场所。在内质网中存在多种调控机制来确保其中的蛋白质被正确地折叠、修饰和组装,以维持内质网稳态,这对于细胞正常的生理活动十分重要。然而,多种物理、化学因素均可使内质网稳态失衡,即在应激条件下,错误折叠和未折叠蛋白质的大量积累将导致内质网胁迫(endoplasmic reticulum stress, ERS),进而会引起未折叠蛋白质响应(unfolded protein response, UPR),极端情况下还会启动细胞程序性死亡(program cell death, PCD)。目前,植物内质网胁迫方面的研究较酵母和动物滞后,因此,从内质网质量控制系统和未折叠蛋白质响应2个方面对植物内质网胁迫现有研究进行了综述,以期为进一步理解内质网胁迫与植物逆境胁迫的关系提供参考。     

蛋白质构象病

蛋白质结构生物学既从蛋白质一级结构序列 ,也从蛋白质空间结构及其动态变化去研究蛋白质的性质和功能。生物医学研究表明蛋白质空间构象发生异常变化会引起疾病发生 ,形成了蛋白质构象病 (Ｐｒｏｔｅｉｎｃｏｎｆｏｒｍａ ｔｉｏｎａｌｄｉｓｅａｓｅｓ)这一新的病理学概念[1] 。1 .蛋白质构象病及其分子构象病理学一般讲 ,引起构象疾病的蛋白质分子与正常蛋白质同时存在于机体内 ,至少部分蛋白质具有正常折叠的空间构象 ,并以正常形态释放。当蛋白质构象异常变化时可导致其生物功能丧失 ,或者引起其后发生的蛋白质聚集与沉积 ,使组织结构…     

未折叠蛋白反应—从内质网到细胞核的细胞内信号传导

在真核细胞中,内质网（ER）中未折叠蛋白聚集时,细胞为生存便会启动未折叠蛋白反应（unfolded protein response,UPR）,这种反应首先发现于酵母中,而其保守性使人们对哺乳动物细胞的RPR有了一定的认识。近年来发现哺乳动物细胞的RPR不仅参与蛋白质合成和分泌通路的调节,还与蛋白质翻译水平下调、细胞周期停滞、细胞凋亡及内质网相关性蛋白质降解（ER-associated degradation,ERAD）等生理过程有关。     

错误折叠蛋白质的聚集效应及其对策

错误折叠蛋白质大量聚集将引发蛋白质构象紊乱症(protein conformational disorder,PCD)。目前的研究发现,蛋白质聚集过程中的中间体(纤维前体,pre—fibfillar)导致细胞膜结构受损,从而诱发细胞凋亡。根据这一原理设计出的抗纤维前体抗体和干扰肽可以作为PCD治疗的一般性方法。此外,该就消除错误折叠蛋白质聚集的研究方向作了展望。     

重组蛋白包涵体的复性研究

重组蛋白在大肠杆菌中的高表达往往形成不可溶、无生物活性的包涵体,需经过变性溶解后,在适当条件下复性形成天然的构象,才可恢复其生物活性．变复性实验是建立在对蛋白质体外折叠机制的了解的基础上．根据近年来对蛋白质折叠机制的认识和重组蛋白包涵体在复性方面的主要进展,论述以下3个方面的内容：1)蛋白质在细胞内的折叠机制;2)蛋白质体外折叠机制;3)蛋白质复性的策略和方法．     

蛋白质错误折叠循环扩增技术

目前发现有多种人类及动物疾病是由体内蛋白质的错误折叠引起的,其中朊毒粒病因具有传染性而备受关注.朊毒粒病研究的核心问题之一是正常细胞朊蛋白（PrPc）向异常致病朊毒粒（PrPsc）转变的机制.蛋白质错误折叠循环扩增技术（protein misfolding cyclic amplification,PMCA）就是最新发明的在体外诱导朊蛋白（PrPc）产生错误折叠生成朊毒粒（prpsc）的技术.该文将概要介绍此项技术的原理、技术要点及在诊断与基础研究方面的应用前景.     

未折叠蛋白质应答

内质网是真核细胞中蛋白质合成、折叠与分泌的重要细胞器.细胞进化出一套完整的机制来监督和帮助内质网内蛋白质的折叠与修饰.而当错误折叠的蛋白质累积时,细胞通过一系列信号转导途径,对其进行应答,包括增强蛋白质折叠能力、停滞大多数蛋白质的翻译、加速蛋白质的降解等.如果内质网功能素乱持续,细胞将最终启动凋亡程序.这些反应被统称为未折叠蛋白质应答(unfolded protein response,UPR).UPR是多个信号转导通路的总称,包括IRE1-XBP1、PERK-ATF4以及ATF6等信号途径.除了应激条件外,UPR还被用于正常生理条件下的调节,例如胆固醇合成代谢的负反馈调控.     

Modulation of neurodegeneration by molecular chaperones

Many neurodegenerative disorders are characterized by conformational changes in proteins that result in misfolding, aggregation and intra- or extra-neuronal accumulation of amyloid fibrils. Molecular chaperones provide a first line of defence against misfolded, aggregation-prone proteins and are among the most potent suppressors of neurodegeneration known for animal models of human disease. Recent studies have investigated the role of molecular chaperones in amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson's disease and polyglutamine diseases. We propose that molecular chaperones are neuroprotective because of their ability to modulate the earliest aberrant protein interactions that trigger pathogenic cascades. A detailed understanding of the molecular basis of chaperone-mediated protection against neurodegeneration might lead to the development of therapies for neurodegenerative disorders that are associated with protein misfolding and aggregation.     

Endoplasmic reticulum proteostasis impairment in aging

Perturbed neuronal proteostasis is a salient feature shared by both aging and protein misfolding disorders. The proteostasis network controls the health of the proteome by integrating pathways involved in protein synthesis, folding, trafficking, secretion, and their degradation. A reduction in the buffering capacity of the proteostasis network during aging may increase the risk to undergo neurodegeneration by enhancing the accumulation of misfolded proteins. As almost one‐third of the proteome is synthetized at the endoplasmic reticulum (ER), maintenance of its proper function is fundamental to sustain neuronal function. In fact, ER stress is a common feature of most neurodegenerative diseases. The unfolded protein response (UPR) operates as central player to maintain ER homeostasis or the induction of cell death of chronically damaged cells. Here, we discuss recent evidence placing ER stress as a driver of brain aging, and the emerging impact of neuronal UPR in controlling global proteostasis at the whole organismal level. Finally, we discuss possible therapeutic interventions to improve proteostasis and prevent pathological brain aging.     

Quality control system of the endoplasmic reticulum and related diseases

The quality control (QC) system of the endoplasmic reticulum (ER) is an important monitoringmechanism in the protein maturation process,which ensures export of properly folded proteins from the ER.Incorrectly or incompletely folded proteins are retained in the ER for refolding or degradation by the ER-residing proteasome.The calnexin/calreticulin cycle and ER-associated degradation are the key elements inQC.These two mechanisms work together to allow incorrectly folded proteins have additional opportunitiesto achieve their native conformations.The QC dysfunction is involved in many diseases caused by mutantproteins,many of which are causes of neurodegenerative disorders.A better understanding of molecularregulation in the QC system will uncover the molecular pathogenic mechanisms of many diseases caused byprotein misfolding and help discover novel strategies for preventing or treating these diseases.     

Quality control of the proteins associated with neurodegenerative diseases

Most neurodegenerative diseases including Alzheimer's disease, Parkinson's disease, Huntington's disease and other polyglutamine diseases are associated with degeneration and death of specific neuronal populations due to misfolding or aggregation of certain proteins. These aggregates often contain ubiquitin that is the signal for proteolysis by the ubiquitin-proteasome system, and chaperone proteins that are involved in the assistance of protein folding. Here we review the role of protein quality control systems in the pathogenesis of neurodegenerative diseases, and aim to learn more from the cooperation between molecular chaperones and ubiquitin-proteasome system responding to cellular protein aggregates, in order to find molecular targets for therapeutic intervention.     

Protein-misfolding diseases and chaperone-based therapeutic approaches

A large number of neurodegenerative diseases in humans result from protein misfolding and aggregation. Protein misfolding is believed to be the primary cause of Alzheimer's disease, Parkinson's disease, Huntington's disease, Creutzfeldt-Jakob disease, cystic fibrosis, Gaucher's disease and many other degenerative and neurodegenerative disorders. Cellular molecular chaperones, which are ubiquitous, stress-induced proteins, and newly found chemical and pharmacological chaperones have been found to be effective in preventing misfolding of different disease-causing proteins, essentially reducing the severity of several neurodegenerative disorders and many other protein-misfolding diseases. In this review, we discuss the probable mechanisms of several protein-misfolding diseases in humans, as well as therapeutic approaches for countering them. The role of molecular, chemical and pharmacological chaperones in suppressing the effect of protein misfolding-induced consequences in humans is explained in detail. Functional aspects of the different types of chaperones suggest their uses as potential therapeutic agents against different types of degenerative diseases, including neurodegenerative disorders.     

Protein Misfolding and Aggregation in Ageing and Disease

Although intensively researched, the fundamental mechanism of protein misfolding that leads to protein aggregation and associated diseases remains somewhat enigmatic. The failure of a protein to correctly fold de novo or to remain correctly folded can have profound consequences on a living system especially when the cellular quality control processes fail to eliminate the rogue proteins. Over 20 different human diseases have now been designated as ‘conformational diseases’ and include neurodegenerative diseases such as Alzheimer’s disease (AD), Huntington’s disease (HD) and Creutzfeldt Jakob disease (CJD) that are becoming increasingly prevalent in an ageing human population. Such diseases are usually characterised by the deposition of specific misfolded proteins as amyloid fibrils and hence are often referred to as the amyloidoses.     

Protein misfolding in the late‐onset neurodegenerative diseases: Common themes and the unique case of amyotrophic lateral sclerosis

Enormous strides have been made in the last 100 years to extend human life expectancy and to combat the major infectious diseases. Today, the major challenges for medical science are age‐related diseases, including cancer, heart disease, lung disease, renal disease, and late‐onset neurodegenerative disease. Of these, only the neurodegenerative diseases represent a class of disease so poorly understood that no general strategies for prevention or treatment exist. These diseases, which include Alzheimer's disease, Parkinson's disease, Huntington's disease, the transmissible spongiform encephalopathies, and amyotrophic lateral sclerosis (ALS), are generally fatal and incurable. The first section of this review summarizes the diversity and common features of the late‐onset neurodegenerative diseases, with a particular focus on protein misfolding and aggregation—a recurring theme in the molecular pathology. The second section focuses on the particular case of ALS, a late‐onset neurodegenerative disease characterized by the death of central nervous system motor neurons, leading to paralysis and patient death. Of the 10% of ALS cases that show familial inheritance (familial ALS), the largest subset is caused by mutations in the SOD1 gene, encoding the Cu, Zn superoxide dismutase (SOD1). The unusual kinetic stability of SOD1 has provided a unique opportunity for detailed structural characterization of conformational states potentially involved in SOD1‐associated ALS. This review discusses past studies exploring the stability, folding, and misfolding behavior of SOD1, as well as the therapeutic possibilities of using detailed knowledge of misfolding pathways to target the molecular mechanisms underlying ALS and other neurodegenerative diseases. Proteins 2013; 81:1285–1303. © 2013 Wiley Periodicals, Inc.     

Protein misfolding and disease; protein refolding and therapy.

Diverse human disorders, including several neurodegenerative diseases and systemic amyloidosis, are thought to arise from the misfolding and aggregation of an underlying protein. Recent findings strongly support this hypothesis and have increased our understanding of the molecular mechanism of protein conformational disorders. Many questions are still pending, but the data overall suggest that correction of protein misfolding constitutes a viable therapeutic strategy for conformational diseases.     

Interplay between protein homeostasis networks in protein aggregation and proteotoxicity

The misfolding and aggregation of disease proteins is characteristic of numerous neurodegenerative diseases. Particular neuronal populations are more vulnerable to proteotoxicity while others are more apt to tolerate the misfolding and aggregation of disease proteins. Thus, the cellular environment must play a significant role in determining whether disease proteins are converted into toxic or benign forms. The endomembrane network of eukaryotes divides the cell into different subcellular compartments that possess distinct sets of molecular chaperones and protein interaction networks. Chaperones act as agonists and antagonists of disease protein aggregation to prevent the accumulation of toxic intermediates in the aggregation pathway. Interacting partners can also modulate the conformation and localization of disease proteins and thereby influence proteotoxicity. Thus, interplay between these protein homeostasis network components can modulate the self‐association of disease proteins and determine whether they elicit a toxic or benign outcome. © 2009 Wiley Periodicals, Inc. Biopolymers 93: 229–236, 2010. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com     

Contribution of glutamatergic signaling to nitrosative stress-induced protein misfolding in normal brain aging and neurodegenerative diseases

Glutamatergic hyperactivity, associated with Ca2+ influx and consequent production of nitric oxide (NO), is potentially involved in both normal brain aging and age-related neurodegenerative disorders. Many neurodegenerative diseases are characterized by conformational changes in proteins that result in their misfolding and aggregation. Normal protein degradation by the ubiquitin-proteasome system can prevent accumulation of aberrantly folded proteins. Our recent studies have linked nitrosative stress to protein misfolding and neuronal cell death. In particular, molecular chaperones - such as protein disulfide isomerase, glucose regulated protein 78, and heat shock proteins - can provide neuroprotection from misfolded proteins by facilitating proper folding and thus preventing aggregation. Here, we present evidence for the hypothesis that NO contributes to normal brain aging and degenerative conditions by S-nitrosylating specific chaperones that would otherwise prevent accumulation of misfolded proteins.