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Biological Calorimetry: Old Friend, New Insights 生物量热法:老朋友,新见解
Pub Date : 2023-01-20 DOI: 10.3390/biophysica3010002
O. Abián, S. Vega, A. Velázquez‐Campoy
Calorimetry is an old experimental technique (first instrument developed in S. XVIII), but it is broadly used and still provides key information for understanding biological processes at the molecular level, particularly, cooperative phenomena in protein interactions. Here, we review and highlight some key aspects of biological calorimetry. Several biological systems will be described in which calorimetry was instrumental for modeling the behavior of the protein and obtaining further biological insight.
量热法是一种古老的实验技术(第一种仪器是在S. XVIII开发的),但它被广泛使用,并且仍然为理解分子水平上的生物过程,特别是蛋白质相互作用中的合作现象提供关键信息。在这里,我们回顾并强调了生物量热法的一些关键方面。将描述几种生物系统,其中量热法用于模拟蛋白质的行为并获得进一步的生物学见解。
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引用次数: 1
A Review of Fifteen Years Developing Computational Tools to Study Protein Aggregation 蛋白质聚集研究计算工具开发15年回顾
Pub Date : 2023-01-18 DOI: 10.3390/biophysica3010001
Carlos Pintado-Grima, O. Bárcenas, Andrea Bartolomé-Nafría, Marc Fornt-Suñé, Valentín Iglesias, J. García-Pardo, S. Ventura
The presence of insoluble protein deposits in tissues and organs is a hallmark of many human pathologies. In addition, the formation of protein aggregates is considered one of the main bottlenecks to producing protein-based therapeutics. Thus, there is a high interest in rationalizing and predicting protein aggregation. For almost two decades, our laboratory has been working to provide solutions for these needs. We have traditionally combined the core tenets of both bioinformatics and wet lab biophysics to develop algorithms and databases to study protein aggregation and its functional implications. Here, we review the computational toolbox developed by our lab, including programs for identifying sequential or structural aggregation-prone regions at the individual protein and proteome levels, engineering protein solubility, finding and evaluating prion-like domains, studying disorder-to-order protein transitions, or categorizing non-conventional amyloid regions of polar nature, among others. In perspective, the succession of the tools we describe illustrates how our understanding of the protein aggregation phenomenon has evolved over the last fifteen years.
组织和器官中不溶性蛋白质沉积物的存在是许多人类疾病的标志。此外,蛋白质聚集体的形成被认为是生产基于蛋白质的治疗方法的主要瓶颈之一。因此,人们对合理化和预测蛋白质聚集非常感兴趣。近二十年来,我们的实验室一直致力于为这些需求提供解决方案。传统上,我们将生物信息学和湿实验室生物物理的核心原理相结合,开发算法和数据库来研究蛋白质聚集及其功能含义。在这里,我们回顾了我们实验室开发的计算工具箱,包括在单个蛋白质和蛋白质组水平上识别序列或结构易聚集区域的程序,工程蛋白质溶解度,发现和评估朊病毒样结构域,研究无序到有序的蛋白质转变,或对极性的非传统淀粉样蛋白区域进行分类,等等。从长远来看,我们描述的一系列工具说明了我们对蛋白质聚集现象的理解在过去十五年中是如何演变的。
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引用次数: 3
Optical Tweezers to Force Information out of Biological and Synthetic Systems One Molecule at a Time 光学镊子一次一个分子地将信息从生物和合成系统中挤出
Pub Date : 2022-12-09 DOI: 10.3390/biophysica2040047
R. Bocanegra, María Ortiz-Rodríguez, Ismael Plaza Garcia-Abadillo, Carlos R-Pulido, B. Ibarra
Over the last few decades, in vitro single-molecule manipulation techniques have enabled the use of force and displacement as controlled variables in biochemistry. Measuring the effect of mechanical force on the real-time kinetics of a biological process gives us access to the rates, equilibrium constants and free-energy landscapes of the mechanical steps of the reaction; this information is not accessible by ensemble assays. Optical tweezers are the current method of choice in single-molecule manipulation due to their versatility, high force and spatial and temporal resolutions. The aim of this review is to describe the contributions of our lab in the single-molecule manipulation field. We present here several optical tweezers assays refined in our laboratory to probe the dynamics and mechano-chemical properties of biological molecular motors and synthetic molecular devices at the single-molecule level.
在过去的几十年里,体外单分子操作技术已经使力和位移成为生物化学中的控制变量。通过测量机械力对生物过程实时动力学的影响,我们可以了解反应的机械步骤的速率、平衡常数和自由能景观;该信息不能通过集合分析来访问。光镊子由于其多功能性、高作用力以及空间和时间分辨率,是目前单分子操作的首选方法。这篇综述的目的是描述我们实验室在单分子操作领域的贡献。我们在这里介绍了在我们的实验室中改进的几种光镊子分析,以在单分子水平上探测生物分子马达和合成分子器件的动力学和机械化学性质。
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引用次数: 0
RBL-2H3 Mast Cell Receptor Dynamics in the Immunological Synapse. RBL-2H3肥大细胞受体在免疫突触中的动态。
Pub Date : 2022-12-01 DOI: 10.3390/biophysica2040038
Ming Chih Tsai, Kathrin Spendier

The RBL-2H3 mast cell immunological synapse dynamics is often simulated with reaction-diffusion and Fokker-Planck equations. The equations focus on how the cell synapse captures receptors following an immune response, where the receptor capture at the immunological site appears to be a delayed process. This article investigates the physical nature and mathematics behind such time-dependent delays. Using signal processing methods, convolution and cross-correlation-type delay capture simulations give a χ-squared range of 22 to 60, in good agreement with experimental results. The cell polarization event is offered as a possible explanation for these capture delays, where polarizing rates measure how fast the cell polarization event occurs. In the case of RBL-2H3 mast cells, polarization appears to be associated with cytoskeletal rearrangement; thus, both cytoskeletal and diffusional components are considered. From these simulations, a maximum polarizing rate ranging from 0.0057 s-2 to 0.031 s-2 is obtained. These results indicate that RBL-2H3 mast cells possess both temporal and spatial memory, and cell polarization is possibly linked to a Turing-type pattern formation.

RBL-2H3肥大细胞免疫突触动力学通常用反应扩散方程和Fokker-Planck方程来模拟。这些方程关注的是细胞突触如何在免疫反应后捕获受体,其中免疫部位的受体捕获似乎是一个延迟的过程。本文研究了这种时间相关延迟背后的物理性质和数学。利用信号处理方法,卷积和互相关型延迟捕获仿真得到的χ-平方范围为22 ~ 60,与实验结果吻合良好。细胞极化事件被提供作为这些捕获延迟的可能解释,其中极化率测量细胞极化事件发生的速度。在RBL-2H3肥大细胞中,极化似乎与细胞骨架重排有关;因此,细胞骨架和扩散成分都被考虑在内。从这些模拟中,得到了最大偏振率范围为0.0057 ~ 0.031 s-2。这些结果表明RBL-2H3肥大细胞具有时间和空间记忆,细胞极化可能与图灵模式的形成有关。
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引用次数: 0
The Forces behind Directed Cell Migration 细胞定向迁移背后的力量
Pub Date : 2022-12-01 DOI: 10.3390/biophysica2040046
Isabela C. Fortunato, R. Sunyer
Directed cell migration is an essential building block of life, present when an embryo develops, a dendritic cell migrates toward a lymphatic vessel, or a fibrotic organ fails to restore its normal parenchyma. Directed cell migration is often guided by spatial gradients in a physicochemical property of the cell microenvironment, such as a gradient in chemical factors dissolved in the medium or a gradient in the mechanical properties of the substrate. Single cells and tissues sense these gradients, establish a back-to-front polarity, and coordinate the migration machinery accordingly. Central to these steps we find physical forces. In some cases, these forces are integrated into the gradient sensing mechanism. Other times, they transmit information through cells and tissues to coordinate a collective response. At any time, they participate in the cellular migratory system. In this review, we explore the role of physical forces in gradient sensing, polarization, and coordinating movement from single cells to multicellular collectives. We use the framework proposed by the molecular clutch model and explore to what extent asymmetries in the different elements of the clutch can lead to directional migration.
定向细胞迁移是生命的重要组成部分,当胚胎发育、树突细胞向淋巴管迁移或纤维化器官无法恢复正常薄壁组织时,就会出现定向细胞迁移。定向细胞迁移通常由细胞微环境的物理化学性质的空间梯度引导,例如溶解在介质中的化学因子的梯度或基质的机械性质的梯度。单个细胞和组织感知这些梯度,建立前后极性,并相应地协调迁移机制。这些步骤的核心是物理力。在某些情况下,这些力被集成到梯度感测机制中。其他时候,它们通过细胞和组织传递信息,以协调集体反应。在任何时候,它们都参与细胞迁移系统。在这篇综述中,我们探讨了物理力在梯度传感、极化和从单细胞到多细胞集体的协调运动中的作用。我们使用分子离合器模型提出的框架,探索离合器不同元件的不对称性在多大程度上会导致定向迁移。
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引用次数: 4
Plasmonic Biosensors Based on Deformed Graphene 基于变形石墨烯的等离子体生物传感器
Pub Date : 2022-11-29 DOI: 10.3390/biophysica2040045
V. Faramarzi, M. Heidari, Nik Humaidi bin Nik Zulkarnine, M. T. Hwang
Rapid, accurate, and label-free detection of biomolecules and chemical substances remains a challenge in healthcare. Optical biosensors have been considered as biomedical diagnostic tools required in numerous areas including the detection of viruses, food monitoring, diagnosing pollutants in the environment, global personalized medicine, and molecular diagnostics. In particular, the broadly emerging and promising technique of surface plasmon resonance has established to provide real-time and label-free detection when used in biosensing applications in a highly sensitive, specific, and cost-effective manner with small footprint platform. In this study we propose a novel plasmonic biosensor based on biaxially crumpled graphene structures, wherein plasmon resonances in graphene are utilized to detect variations in the refractive index of the sample medium. Shifts in the resonance wavelength of the plasmon modes for a given change in the RI of the surrounding analyte are calculated by investigating the optical response of crumpled graphene structures on different substrates using theoretical computations based on the finite element method combined with the semiclassical Drude model. The results reveal a high sensitivity of 4990 nm/RIU, corresponding to a large figure-of-merit of 20 for biaxially crumpled graphene structures on polystyrene substrates. We demonstrate that biaxially crumpled graphene exhibits superior sensing performance compared with a uniaxial structure. According to the results, crumpled graphene structures on a titanium oxide substrate can improve the sensor sensitivity by avoiding the damping effects of polydimethylsiloxane substrates. The enhanced sensitivity and broadband mechanical tunability of the biaxially crumpled graphene render it a promising platform for biosensing applications.
生物分子和化学物质的快速、准确和无标签检测仍然是医疗保健领域的一项挑战。光学生物传感器被认为是许多领域所需的生物医学诊断工具,包括病毒检测、食品监测、环境污染物诊断、全球个性化医学和分子诊断。特别是,表面等离子体共振这一新兴且有前景的技术已经建立起来,当以高灵敏度、特异性和成本效益的方式在小占地面积平台的生物传感应用中使用时,可以提供实时和无标记的检测。在这项研究中,我们提出了一种基于双轴褶皱石墨烯结构的新型等离子体生物传感器,其中石墨烯中的等离子体共振用于检测样品介质折射率的变化。通过使用基于有限元方法和半经典Drude模型的理论计算研究不同衬底上皱缩石墨烯结构的光学响应,计算了周围分析物的RI发生给定变化时等离子体模式的共振波长的偏移。结果显示了4990nm/RIU的高灵敏度,对应于聚苯乙烯衬底上双轴皱缩石墨烯结构的20的大优值。我们证明,与单轴结构相比,双轴皱缩石墨烯表现出优异的传感性能。结果表明,氧化钛衬底上的石墨烯结构可以通过避免聚二甲基硅氧烷衬底的阻尼效应来提高传感器的灵敏度。双轴皱缩石墨烯增强的灵敏度和宽带机械可调谐性使其成为生物传感应用的一个有前途的平台。
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引用次数: 4
Adherent Moving of Polymers in Spherical Confined Binary Semiflexible Ring Polymer Mixtures 聚合物在球形约束二元半柔性环聚合物混合物中的粘附运动
Pub Date : 2022-11-24 DOI: 10.3390/biophysica2040044
Xiaolin Zhou, Wan Wei
Based on the coarse-grained model, we used molecular dynamics methods to calculate and simulate a semiflexible long ring–semiflexible short ring blended polymer system confined in a hard sphere. We systematically studied the distribution and motion characteristics of the long ring chain. The results show that when the short ring is short enough (Lshort < 20), the long ring (Llong = 50) is separated from the blend system and then distributed against the inner wall. As the length of the short ring increases (Lshort ≥ 20), the long ring can no longer be separated from the blending system. Moreover, we found that the long ring demonstrates a random direction of adherent walking behavior on the inner surface of the hard sphere. The velocity of the long ring decreases with the increase in the short ring length Lshort. Specifically for Lshort ≥ 20, the system does not undergo phase separation and the speed of the long ring decreases sharply along with the long ring distributed inside the confined bulk. This is related to the inner wall layer moving faster than the inside bulk of the restricted system. Our simulation results can help us to understand the distribution of macromolecules in biological systems in confined systems, including the restricted chromosome partitioning distribution and packing structure of circular DNA molecules.
基于粗粒模型,我们使用分子动力学方法计算和模拟了一个半柔性长环-半柔性短环混合聚合物系统,该系统被限制在一个硬球中。我们系统地研究了长环链的分布和运动特性。结果表明,当短环足够短(Lshort<20)时,长环(Llong=50)从共混体系中分离出来,然后靠着内壁分布。随着短环长度的增加(Lshort≥20),长环不能再与共混体系分离。此外,我们发现长环在硬球的内表面上表现出粘附行走行为的随机方向。长环的速度随着短环长度Lshort的增加而减小。特别是当Lshort≥20时,系统不发生相分离,长环的速度随着分布在受限体积内的长环而急剧下降。这与内壁层比受限制系统的内部体积移动得更快有关。我们的模拟结果可以帮助我们了解生物系统中大分子在受限系统中的分布,包括环状DNA分子的受限染色体分配分布和堆积结构。
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引用次数: 0
Characterisation of Amyloid Aggregation and Inhibition by Diffusion-Based Single-Molecule Fluorescence Techniques 基于扩散的单分子荧光技术表征淀粉样蛋白聚集和抑制
Pub Date : 2022-11-21 DOI: 10.3390/biophysica2040043
David Polanco, Alejandra Carrancho, P. Gracia, N. Cremades
Protein amyloid aggregation has been associated with more than 50 human disorders, including the most common neurodegenerative disorders Alzheimer’s and Parkinson’s disease. Interfering with this process is considered as a promising therapeutic strategy for these diseases. Our understanding of the process of amyloid aggregation and its role in disease has typically been limited by the use of ensemble-based biochemical and biophysical techniques, owing to the intrinsic heterogeneity and complexity of the process. Single-molecule techniques, and particularly diffusion-based single-molecule fluorescence approaches, have been instrumental to obtain meaningful information on the dynamic nature of the fibril-forming process, as well as the characterisation of the heterogeneity of the amyloid aggregates and the understanding of the molecular basis of inhibition of a number of molecules with therapeutic interest. In this article, we reviewed some recent contributions on the characterisation of the amyloid aggregation process, the identification of distinct structural groups of aggregates in homotypic or heterotypic aggregation, as well as on the study of the interaction of amyloid aggregates with other molecules, allowing the estimation of the binding sites, affinities, and avidities as examples of the type of relevant information we can obtain about these processes using these techniques.
淀粉样蛋白聚集与50多种人类疾病有关,包括最常见的神经退行性疾病阿尔茨海默氏症和帕金森病。干扰这一过程被认为是治疗这些疾病的一种有希望的治疗策略。由于淀粉样蛋白聚集过程的内在异质性和复杂性,我们对淀粉样蛋白聚集过程及其在疾病中的作用的理解通常受到基于集合的生化和生物物理技术的限制。单分子技术,特别是基于扩散的单分子荧光方法,有助于获得有关原纤维形成过程的动态性质的有意义的信息,以及淀粉样蛋白聚集体异质性的特征,以及对一些具有治疗意义的分子抑制的分子基础的理解。在这篇文章中,我们回顾了最近在淀粉样蛋白聚集过程的表征,在同型或异型聚集中不同结构群的识别,以及淀粉样蛋白聚集与其他分子相互作用的研究方面的一些贡献,允许估计结合位点,亲和力和亲和力,作为我们可以使用这些技术获得这些过程相关信息类型的例子。
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引用次数: 0
Birth and Early Steps of the Organization of Biophysics in Spain 西班牙生物物理学组织的诞生和早期步骤
Pub Date : 2022-11-19 DOI: 10.3390/biophysica2040042
F. Goñi
In the 1960s, Biophysics was an unheard of scientific field in Spain, and even outside Spain, the distinction between Biophysics and Molecular Biology was not clear at the time. This paper describes briefly the developments that led to the foundation of the Spanish National Committee for Biophysics (1981) and of the Spanish Biophysical Society (1987), the incorporation of Spain into IUPAB and EBSA, and the normalized presence of Biophysics as a compulsory subject in undergraduate curricula in Spain.
在20世纪60年代,生物物理学在西班牙是一个闻所未闻的科学领域,即使在西班牙以外,生物物理学和分子生物学之间的区别当时也不清楚。本文简要介绍了西班牙国家生物物理委员会(1981年)和西班牙生物物理学会(1987年)的成立、西班牙加入IUPAB和EBSA以及生物物理作为西班牙本科必修课的规范化发展。
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引用次数: 0
Low-Energy Electron Generation for Biomolecular Damage Inquiry: Instrumentation and Methods 低能电子产生用于生物分子损伤研究:仪器和方法
Pub Date : 2022-11-17 DOI: 10.3390/biophysica2040041
E. Alizadeh, Dipayan Chakraborty, S. Ptasińska
Technological advancement has produced a variety of instruments and methods to generate electron beams that have greatly assisted in the extensive theoretical and experimental efforts devoted to investigating the effect of secondary electrons with energies approximately less than 100 eV, which are referred as low-energy electrons (LEEs). In the past two decades, LEE studies have focused on biomolecular systems, which mainly consist of DNA and proteins and their constituents as primary cellular targets of ionizing radiation. These studies have revealed that compared to other reactive species produced by high-energy radiation, LEEs have distinctive pathways and considerable efficiency in inducing lethal DNA lesions. The present work aims to briefly discuss the current state of LEE production technology and to motivate further studies and improvements of LEE generation techniques in relation to biological electron-driven processes associated with such medical applications as radiation therapy and cancer treatment.
技术进步产生了各种产生电子束的仪器和方法,这些仪器和方法极大地帮助了研究能量约小于100eV的二次电子(称为低能电子)效应的广泛理论和实验工作。在过去的二十年里,LEE的研究主要集中在生物分子系统上,该系统主要由DNA和蛋白质及其成分组成,是电离辐射的主要细胞靶点。这些研究表明,与高能辐射产生的其他反应物种相比,LEE在诱导致命DNA损伤方面具有独特的途径和相当高的效率。本工作旨在简要讨论LEE生产技术的现状,并推动进一步研究和改进与放射治疗和癌症治疗等医学应用相关的生物电驱动过程中的LEE生成技术。
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引用次数: 3
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Biophysica
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