分子生物物理学方法结构、动力学、功能

分子生物物理学旨在应用物理学方法了解生物大分子的结构及其相互作用，利用经典热力学及近期发展的各种物理学方法来检测和操控生物分子。这些方法包括质谱、流体力学、电子显微学、衍射和晶体学、分子动力学模拟以及核磁共振，它们相辅相成，各有其特定的优势与适用范围。     

芳华人生"显微"科技"半边天"--记中国科学院院士李方华

李方华。著名物理学家、电子显微学家。中国单晶体电子衍射结构分析的开创者。中国建立并发展高分辨电子显微学的代表人物。1932年1月出生于香港。1956年毕业进入中国科学院物理研究所。工作至今。曾任中国电子显微学会理事长。中国物理学会理事。     

植物细胞内膜运输调控机制及相关前沿电子显微镜技术应用的研究进展

真核细胞内的蛋白质等生物大分子通常需要借助内膜系统才能被运送到目标部位并发挥相应功能.植物细胞内膜系统由核膜、细胞膜以及多个膜包裹的细胞器组成,包括内质网、高尔基体、反式高尔基体网络、液泡前体/多囊泡体、液泡和自噬体等.植物蛋白质在内膜系统中可以通过囊泡进行运输,对生物个体的生长发育和环境应答至关重要.本文系统地介绍了植物细胞内囊泡介导下多种蛋白运输途径的调控机制研究进展.此外,鉴于前沿电子显微镜技术在推动本领域研究发展中发挥的巨大贡献,重点阐述冷冻电子显微镜、电子断层扫描技术、冷冻聚焦离子束和光电联用技术在植物细胞内膜运输研究中的应用.最后,对本领域研究现况进行了总结,并提出有待解决的关键问题和对未来发展的展望.     

生物大分子纯化系统的应用与管理

生物大分子纯化系统是一类利用色谱技术,对各种生物大分子进行分离纯化的自动化运行系统。该文阐述了凝胶色谱柱的种类、生物大分子的分离纯化方法、纯化工艺中不同色谱技术的衔接及如何依据蛋白质等生物大分子的理化性质合理设计色谱方案、生物大分子纯化系统的维护与保养,并简要概述了生物大分子纯化系统在生命科学研究中的应用。     

质谱和核磁共振技术在生物大分子研究中的应用

介绍了研究生物大分子的意义和“电喷雾”质谱技术、“软激光解吸”质谱技术以及核磁共振技术在生物大分子中的应用与进展.     

荧光探针在生物大分子中的应用

介绍了荧光探针定量测定生物大分子的基本原理、测定方法及新型荧光探针技术在生物大分子研究中的一些新进展.     

生物大分子电化学传感器的研究进展

生物大分子电化学传感器是围绕生物大分子的检测分析研究而开发出来的一系列电化学传感器,在这些传感器中尤其是以纳米间隙电化学传感器性能最为优异,在实际的检测研究中表现出超高灵敏度、微型化、较好的选择性、所需检测样品少、检测速度快、非常便捷等明显优势,因而受到了研究人员的极大关注.本文简单介绍了生物大分子电化学传感器的原理、分类、加工技术及特点,着重分析了间隙电化学传感器较比常规电化学传感器所具备的优越特点.综述了近十年来生物大分子的检测研究进展,特别是纳米间隙电化学传感器在生物大分子检测研究中的应用,并对生物大分子电化学传感器的研究和开发进行展望.     

电子显微镜在本科实验教学中的应用

电子显微镜是广泛应用于生物科学、材料科学、纳米科学等多个领域的大型仪器。对透射电子显微镜和扫描电子显微镜应用于中山大学本科专业选修课"现代生物电子显微学"和核心通识课"电镜下的微观世界"中的实践进行了讨论。通过教学改革的探索,激发了学生的求知欲,上课积极性提高,主动性增强,电子显微镜实验教学效果得到了有效的提升。     

生物质谱的研究及其应用

发展生物质谱是为解决生命科学中有关生物物质分析问题,主要借助软电离质谱技术对生物大分子如蛋白质、核酸和多糖等进行结构分析.生物质谱已成为质谱学中最活跃的研究领域,推动了质谱分析理论和技术的发展.笔者简要介绍了生物质谱的发展,重点讨论因"发明了对生物大分子的质谱分析法"而荣获2002年诺贝尔化学奖金一半的美国科学家约翰.芬恩教授(John B.Fenn)和日本科学家田中耕一工程师(Koichi Tanaka)分别研发的电喷雾电离质谱(ESI-MS)和基质辅助激光解吸电离质谱(MALDI-MS),特别阐述它们应用于蛋白质、核酸和糖结构分析及基因组学、蛋白质组学、糖组学等生物组学和化学组学中.     

分子构象—电子相互作用的Green函数理论

本文将 Green 函数理论推广于研究分子构象和电子(以及电磁场)的相互作用,给出了一套适宜于处理生物大分子构象变化的理论形式。     

异军突起的冷冻电镜技术——2017年度诺贝尔化学奖成果简析

 2017年诺贝尔化学奖授予Jacques Dubochet、Joachim Frank和Richard Henderson 3位科学家,以表彰他们在开发用于溶液中生物分子高分辨率结构测定的冷冻电镜技术方面的贡献。本文将简述这3位科学家的获奖工作,并展望冷冻电镜技术的发展前景。     

Switch-based mechanism of kinesin motors

Kinesin motors are specialized enzymes that use hydrolysis of ATP to generate force and movement along their cellular tracks, the microtubules. Although numerous biochemical and biophysical studies have accumulated much data that link microtubule-assisted ATP hydrolysis to kinesin motion, the structural view of kinesin movement remains unclear. This study of the monomeric kinesin motor KIF1A combines X-ray crystallography and cryo-electron microscopy, and allows analysis of force-generating conformational changes at atomic resolution. The motor is revealed in its two functionally critical states-complexed with ADP and with a non-hydrolysable analogue of ATP. The conformational change observed between the ADP-bound and the ATP-like structures of the KIF1A catalytic core is modular, extends to all kinesins and is similar to the conformational change used by myosin motors and G proteins. Docking of the ADP-bound and ATP-like crystallographic models of KIF1A into the corresponding cryo-electron microscopy maps suggests a rationale for the plus-end directional bias associated with the kinesin catalytic core.     

The molecular sociology of the cell

Proteomic studies have yielded detailed lists of the proteins present in a cell. Comparatively little is known, however, about how these proteins interact and are spatially arranged within the 'functional modules' of the cell: that is, the 'molecular sociology' of the cell. This gap is now being bridged by using emerging experimental techniques, such as mass spectrometry of complexes and single-particle cryo-electron microscopy, to complement traditional biochemical and biophysical methods. With the development of integrative computational methods to exploit the data obtained, such hybrid approaches will uncover the molecular architectures, and perhaps even atomic models, of many protein complexes. With these structures in hand, researchers will be poised to use cryo-electron tomography to view protein complexes in action within cells, providing unprecedented insights into protein-interaction networks.     

冷冻电子显微技术——2017年度诺贝尔化学奖成果简析

 2017年度诺贝尔化学奖授予瑞士洛桑大学的Jacques Dubochet、美国哥伦比亚大学的Joachim Frank和英国MRC分子生物学实验室的Richard Henderson 3位科学家。本文简要介绍冷冻电镜的发展历史、3位诺贝尔奖获得者在冷冻电镜技术发展过程中的贡献以及国际和国内的最新研究进展。     

Molecular structure of F-actin and location of surface binding sites

Comparisons of three-dimensional maps of vertebrate muscle thin filaments obtained by cryo-electron microscopy and image analysis, reveal the molecular structure of F-actin, the location of the C terminus of the monomer and the positions of the binding sites of tropomyosin, the myosin head and the N-terminal portion of the myosin A1 light chain on the filament. These data provide strong constraints for evaluating models built from the atomic structure of the monomer and the subsequent identification of molecular contacts.     

Envelope structure of Semliki Forest virus reconstructed from cryo-electron micrographs

The basic principles of the architecture of many viral protein shells have been successfully established from electron microscopy and X-ray data, but enveloped viruses have been more difficult to study because they resist crystallization and are easily deformed when prepared for electron microscopy. To avoid the limitations of conventional techniques when applied to enveloped viruses, we have used a cryo-electron microscopy method in which unfixed and unstained viruses are observed in an unsupported thin layer of vitrified suspension. Because of electron beam damage, the many different views required for high-resolution three-dimensional reconstruction cannot be obtained from a tilt series of the same particle. The images of many differently oriented viruses are combined using a novel reconstruction method, 'reconstruction by optimized series expansion' (ROSE). The structure of the envelope of Semliki Forest virus has been reconstructed to 3.5-nm resolution. The T = 4 geometry of the surface lattice, the shape of the trimeric spikes and their arrangement on the lipid bilayer are visualized.     

Structure and mechanism of the chromatin remodelling factor ISW1a

Site-specific recognition of DNA in eukaryotic organisms depends on the arrangement of nucleosomes in chromatin. In the yeast Saccharomyces cerevisiae, ISW1a and related chromatin remodelling factors are implicated in establishing the nucleosome repeat during replication and altering nucleosome position to affect gene activity. Here we have solved the crystal structures of S. cerevisiae ISW1a lacking its ATPase domain both alone and with DNA bound at resolutions of 3.25?? and 3.60??, respectively, and we have visualized two different nucleosome-containing remodelling complexes using cryo-electron microscopy. The composite X-ray and electron microscopy structures combined with site-directed photocrosslinking analyses of these complexes suggest that ISW1a uses a dinucleosome substrate for chromatin remodelling. Results from a remodelling assay corroborate the dinucleosome model. We show how a chromatin remodelling factor could set the spacing between two adjacent nucleosomes acting as a 'protein ruler'.     

Structure of the E. coli protein-conducting channel bound to a translating ribosome

Secreted and membrane proteins are translocated across or into cell membranes through a protein-conducting channel (PCC). Here we present a cryo-electron microscopy reconstruction of the Escherichia coli PCC, SecYEG, complexed with the ribosome and a nascent chain containing a signal anchor. This reconstruction shows a messenger RNA, three transfer RNAs, the nascent chain, and detailed features of both a translocating PCC and a second, non-translocating PCC bound to mRNA hairpins. The translocating PCC forms connections with ribosomal RNA hairpins on two sides and ribosomal proteins at the back, leaving a frontal opening. Normal mode-based flexible fitting of the archaeal SecYEbeta structure into the PCC electron microscopy densities favours a front-to-front arrangement of two SecYEG complexes in the PCC, and supports channel formation by the opening of two linked SecY halves during polypeptide translocation. On the basis of our observation in the translocating PCC of two segregated pores with different degrees of access to bulk lipid, we propose a model for co-translational protein translocation.     

The Ndc80 kinetochore complex forms oligomeric arrays along microtubules

The Ndc80 complex is a key site of regulated kinetochore-microtubule attachment (a process required for cell division), but the molecular mechanism underlying its function remains unknown. Here we present a subnanometre-resolution cryo-electron microscopy reconstruction of the human Ndc80 complex bound to microtubules, sufficient for precise docking of crystal structures of the component proteins. We find that the Ndc80 complex binds the microtubule with a tubulin monomer repeat, recognizing α- and β-tubulin at both intra- and inter-tubulin dimer interfaces in a manner that is sensitive to tubulin conformation. Furthermore, Ndc80 complexes self-associate along protofilaments through interactions mediated by the amino-terminal tail of the NDC80 protein, which is the site of phospho-regulation by Aurora B kinase. The complex's mode of interaction with the microtubule and its oligomerization suggest a mechanism by which Aurora B could regulate the stability of load-bearing kinetochore-microtubule attachments.