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Structural insights into the functions of Raf1 and Bsd2 in hexadecameric Rubisco assembly

Molecular Plant 2023年第12期16卷 1927-1936页

作者： Ran Wang Hui Song Wenjuan Zhang Ning Wang Shijia Zhang Ruiqi Shao Cuimin Liu State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental BiologyThe Innovative Academy of Seed DesignChineseAcademyof SciencesBeijing100101China University of Chinese Academy of Sciences Bejing 100101China

Hexadecameric formI Rubisco,which consisting consists of eight large(RbcL)and eight small(RbcS)subunits,is the most abundant enzyme on earth.Extensive efforts to engineer an improved Rubisco to speed up its catalytic efficiency and ultimately increase agricultural productivity.However,difficulties with correct folding and assembly in foreign hosts or in vitro have hampered the genetic manipulation of hexadecameric Rubisco.In this study,we reconstituted Synechococcus sp.Pcc6301 Rubisco in vitro using the chaperonin system and assembly factors from cyanobacteria and Arabidopsis thaliana(At).Rubisco holoenzyme was produced in the presence of cyanobacterial Rubisco accumulation factor 1(Raf1)alone or both AtRaf1 and bundle-sheath defective-2(AtBsd2)from Arabidopsis.RbcL released from GroEL is assembly capable in the presence of ATP,and AtBsd2 functions downstream of AtRaf1.Cryo-EM structures of RbcL8-AtRaf18,RbcL8-AtRaf14-AtBsd2s,and RbcLs revealed that the interactions between RbcL and AtRaf1 are looser than those between prokaryotic RbcL and Raf1,with AtRaf1 tilting 7°farther away from RbcL.AtBsd2 stabilizes the flexible regions of RbcL,including the N and C termini,the 60s loop,and loop 6.Using these data,combined with previous findings,we propose the possible biogenesis pathways of prokaryotic and eukaryotic Rubisco.

关键词： Rubisco chaperone protein folding protein assembly assembly factor

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protein assembly: A New Platform to Develop Biomimetic System

Protein Assembly: A New Platform to Develop Biomimetic Syste...

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第十一届全国化学生物学学术会议

作者：刘俊秋吉林大学

Sophisticated protein self-assemblies have attracted great scientific interests in recent few decades due to their various potential applications.The design and control of proteins into hierarchical nanostructures via self-assembly strategies offers unique advantages in understanding the mechanism of naturally occurring protein assemblies and in creating various functional biomaterials with advanced properties.protein self-assembly into exquisite,complicated yet high-ordered architectures represents the supreme wisdom of nature.However,precisely manipulating protein self-assembling behaviors in vitro is a great challenge.By taking advantage of supramolecular strategies such as the metal ion chelating interactions,host-guest interaction and non-specific protein-protein interactions,accuracy control of the orientation of protein self-assembly has been achieved.The designed nanostructures have been used as biomimetic scaffolds for developing biomimetic enzymes,light harvesting system and muscle mimics.

关键词： protein assembly Biomimetic System Supramolecular chemistry

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Metal induced self-assembly of designed V-shape protein into 2D wavy supramolecular nanostructure

Metal induced self-assembly of designed V-shape protein into...

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第十届全国化学生物学学术会议

作者： Qiao,Shanpeng Lang,Chao Wang,Ruidi Li,Xiumei Yan,Tengfei Pan,Tiezheng Zhao,Linlu Fan,Xiaotong Zhang,Xin Hou,Chunxi Luo,Quan Xu,Jiayun Liu,Junqiu State Key Laboratory of Supramolecular Structure and Materials College of ChemistryJilin University

In order to understand and imitate the more complex bio-processes and fascinating functions in nature, protein self-assembly has been studied and has attracted more and more interest in recent years. Artificial self-assemblies of proteins have been constructed through many strategies.[1-3] However, the design of complicated protein self-assemblies utilizing the special profile of building blocks remains a challenge. We herein report linear and 2D nanostructures constructed from a V shape SMAC protein and induced by metal coordination. Zigzag nanowires and wavy 2D nanostructures have been demonstrated by AFM and TEM. The zigzag nanowires can translate to a 2D nanostructure with an excess of metal ions, which reveals the step by step assembly process. Fluorescence and UV/Vis spectra have also been obtained to further study the mechanism and process of self-assembly. Upon the protein nanostructure, fluorescence resonance energy transfer（FRET） could also be detected using fluorescein modified proteins as building blocks. This article provides an approach for designing and controlling self-assembled protein nanostructures with a distinctive topological morphology.

关键词： protein assembly Metal coordination FRET

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Enzyme-Triggered Defined protein Nanoarrays:Efficient Light-Harvesting Systems to Mimic Chloroplasts

Enzyme-Triggered Defined Protein Nanoarrays:Efficient Light-...

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第十届全国化学生物学学术会议

作者： Linlu Zhao Junqiu Liu State Key Laboratory of Supramolecular Structure and Materials College of ChemistryJilin University

The elegance and efficiency by which chloroplasts harvest solar energy and conduct energy transfer have been a source of inspiration for chemists to mimic such process. However, precise manipulation to obtain orderly arranged antenna chromophores in constructing artificial chloroplast mimics was a great challenge, especially from the structural similarity and bioaffinity standpoints. Here we reported a design strategy that combined covalent and noncovalent interactions to prepare a protein-based light-harvesting system to mimic chloroplasts. Cricoid stable protein one（SP1） was utilized as a building block model. Under enzyme-triggered covalent protein assembly, mutant SP1 with tyrosine（Tyr） residues at the designated sites can couple together to form nanostructures. Through controlling the Tyr sites on the protein surface, we can manipulate the assembly orientation to respectively generate 1D nanotubes and 2D nanosheets. The excellent stability endowed the self-assembled protein architectures with promising applications. We further integrated quantum dots（QDs） possessing optical and electronic properties with the 2D nanosheets to fabricate chloroplast mimics. By attaching different sized QDs as donor and acceptor chromophores to the negatively charged surface of SP1-based protein nanosheets via electrostatic interactions, we successfully developed an artificial light-harvesting system. The assembled protein nanosheets structurally resembled the natural thylakoids, and the QDs can achieve pronounced FRET phenomenon just like the chlorophylls. Therefore, the coassembled system was meaningful to explore the photosynthetic process in vitro, as it was designed to mimic the natural chloroplast.

关键词： protein nanostructures protein assembly Light-harvesting Quantum dot Chloroplast mimic

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Construction of Functional protein Nanowires by Supramolecular Interactions

Construction of Functional Protein Nanowires by Supramolecul...

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第八届全国化学生物学学术会议

作者： Chunxi Hou Shanpeng Qiao Junqiu Liu State Key Laboratory of SupCramolecular Structure and Materials College of Chemistry Jilin University

protein nanostructures are dimensional homogenous or heterogenous protein assemblies such as virus capsids,tubulins and actins with sophisticated physical, chemical and structural properties.In natural,protein nanostr... 详细信息

关键词： protein assembly Host guest interactions protein nanowires Biomaterials

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