bioRxiv - Biophysics最新文献_第7页

Feather keratin in Pavo cristatus: A tentative structure Pavo cristatus 的羽毛角蛋白：暂定结构

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.08.611866

Peter Russ, Helmut O K Kirchner, Herwig Peterlik, Ingrid M Weiss

引用次数: 0

Biophysical modeling of membrane curvature generation and curvature sensing by the glycocalyx 膜曲率生成和糖萼曲率感应的生物物理建模

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.07.611813

Ke Xiao, Sujeong Park, Jeanne Stachowiak, Padmini Rangamani

{"title":"Biophysical modeling of membrane curvature generation and curvature sensing by the glycocalyx","authors":"Ke Xiao, Sujeong Park, Jeanne Stachowiak, Padmini Rangamani","doi":"10.1101/2024.09.07.611813","DOIUrl":"https://doi.org/10.1101/2024.09.07.611813","url":null,"abstract":"Generation of membrane curvature is fundamental to cellular function. Recent studies have established that the glycocalyx, a sugar-rich polymer layer at the cell surface, can generate membrane curvature. While there have been some theoretical efforts to understand the interplay between the glycocalyx and membrane bending, there remain open questions about how the properties of the glycocalyx affect membrane bending. For example, the relationship between membrane curvature and the density of glycosylated proteins on its surface remains unclear. In this work, we use polymer brush theory to develop a detailed biophysical model of the energetic interactions of the glycocalyx with the membrane. Using this model, we identify the conditions under which the glycocalyx can both generate and sense curvature. Our model predicts that the extent of membrane curvature generated depends on the grafting density of the glycocalyx and the length of the polymers constituting the glycocalyx. Furthermore, when coupled with the intrinsic membrane properties such as spontaneous curvature and a line tension along the membrane, the curvature generation properties of the glycocalyx are enhanced. These predictions were tested experimentally by examining the propensity of glycosylated transmembrane proteins to drive the assembly of highly-curved filopodial protrusions at the plasma membrane of adherent mammalian cells. Our model also predicts that the glycocalyx has curvature-sensing capabilities, in agreement with the results of our experiments.\u0000Thus, our study develops a quantitative framework for mapping the properties of the glycocalyx to the curvature generation capability of the membrane.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178221","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Calibration of FRET-based biosensors using multiplexed biosensor barcoding 利用多重生物传感器条形码校准基于 FRET 的生物传感器

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.04.610346

Jhen-Wei Wu, Jr-Ming Yang, Chao-Cheng Chen, Gabriel Au, Suyang Wang, Gia-Wei Chern, Chuan-Hsiang Huang

{"title":"Calibration of FRET-based biosensors using multiplexed biosensor barcoding","authors":"Jhen-Wei Wu, Jr-Ming Yang, Chao-Cheng Chen, Gabriel Au, Suyang Wang, Gia-Wei Chern, Chuan-Hsiang Huang","doi":"10.1101/2024.09.04.610346","DOIUrl":"https://doi.org/10.1101/2024.09.04.610346","url":null,"abstract":"Förster resonance energy transfer (FRET) between fluorescent proteins (FPs) is widely used in the design of genetically encoded fluorescent biosensors, which are powerful tools for monitoring the dynamics of biochemical activities in live cells. FRET ratio, defined as the ratio between acceptor and donor signals, is often used as a proxy for the actual FRET efficiency, which must be corrected for signal crosstalk using donor-only and acceptor-only samples. However, the FRET ratio is highly sensitive to imaging conditions, making direct comparisons across different experiments and over time challenging. Inspired by a method for multiplexed biosensor imaging using barcoded cells, we reasoned that calibration standards with fixed FRET efficiency can be introduced into a subset of cells for normalization of biosensor signals. Our theoretical analysis indicated that the FRET ratio of high-FRET species relative to non-FRET species slightly decreases at high excitation intensity, suggesting the need for calibration using both high and low FRET standards. To test these predictions, we created FRET donor-acceptor pairs locked in \"FRET-ON\" and \"FRET-OFF\" conformations and introduced them into a subset of cells using the cell barcoding strategy. Our results confirmed the theoretical predictions and showed that the calibrated FRET ratio is independent of imaging settings. We also provided a strategy for calculating the FRET efficiency. Together, our study presents a simple strategy for calibrated and highly multiplexed imaging of FRET biosensors, facilitating reliable comparisons across experiments and supporting long-term imaging applications.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178243","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

A highly active bacterial actin actuates the polymerization of another isoform essential for swimming motility of Spiroplasma 一种高活性细菌肌动蛋白促使螺浆虫游泳运动所必需的另一种同工酶聚合

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.04.611326

Daichi Takahashi, Hana Kiyama, Hideaki Matsubayashi, Ikuko Fujiwara, Makoto Miyata

{"title":"A highly active bacterial actin actuates the polymerization of another isoform essential for swimming motility of Spiroplasma","authors":"Daichi Takahashi, Hana Kiyama, Hideaki Matsubayashi, Ikuko Fujiwara, Makoto Miyata","doi":"10.1101/2024.09.04.611326","DOIUrl":"https://doi.org/10.1101/2024.09.04.611326","url":null,"abstract":"Spiroplasma is a wall−less helical bacterium possessing five isoforms of bacterial actin MreBs (SMreB1−5) for its swimming, speculated to be the sole motility system driven by endogenous bacterial actin proteins. Its detailed molecular mechanism remains elusive due to the lack of soluble constructs of SMreB1 essential for Spiroplasma swimming. Here, we isolated soluble SMreB1 of Spiroplasma eriocheiris (SpeMreB1) and evaluated its activity. The phosphate release rate and fold changes of polymerization−critical concentrations over the nucleotide states of SpeMreB1 are the highest among the MreB family proteins. SpeMreB1 interacts with polymerized SpeMreB5, another SMreB essential for Spiroplasma swimming, and decreases SpeMreB5 filament amount depending on the nucleotide state. A decrease in SpeMreB5 filament amount is independent of SpeMreB1 polymerization, although it is essential for swimming motility. SpeMreB1 binds to negatively charged lipids, regardless of their nucleotide state. These results suggest that SpeMreB1 manages SpeMreB5 filaments to drive Spiroplasma swimming.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178251","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Restoration of 3-D structure of insect flight muscle from a rotationally averaged 2-D X-ray diffraction pattern 从旋转平均二维 X 射线衍射图样还原昆虫飞行肌肉的三维结构

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.04.611338

Hiroyuki Iwamoto

{"title":"Restoration of 3-D structure of insect flight muscle from a rotationally averaged 2-D X-ray diffraction pattern","authors":"Hiroyuki Iwamoto","doi":"10.1101/2024.09.04.611338","DOIUrl":"https://doi.org/10.1101/2024.09.04.611338","url":null,"abstract":"The contractile machinery of muscle, especially that of skeletal muscle, has a very regular array of contractile protein filaments, and gives rise to a very complex and informative diffraction pattern when irradiated with X-rays. However, the analysis of the diffraction patterns is often difficult, because (1) only rotationally averaged diffraction patterns can be obtained, resulting in substantial loss of information, and (2) the contractile machinery contains two different sets of protein filaments (actin and myosin) with different helical symmetries, and the reflections originating from them are often overlapped. These problems may be solved if the real-space 3-D structure of the contractile machinery is directly calculated from the diffraction pattern. Here we demonstrate that, by using the conventional phase-retrieval algorithm (hybrid input-output), the real-space 3-D structure of the contractile machinery can be well restored from a single rotationally averaged 2-D diffraction pattern. In this calculation, we used a model structure of insect flight muscle, known to have a very regular structure. Possibilities of extending this technique to the actual muscle diffraction patterns is discussed.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178220","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Direct contact of the bacterial surface induces phase separation in the host phagosome membrane 细菌表面的直接接触导致宿主吞噬体膜相分离

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.04.611260

Aher Jayesh Bhausaheb, Aniruddha N, Debraj Koiri, Jafarulla Shaikh, Sandeep Choubey, Mohammed Saleem

{"title":"Direct contact of the bacterial surface induces phase separation in the host phagosome membrane","authors":"Aher Jayesh Bhausaheb, Aniruddha N, Debraj Koiri, Jafarulla Shaikh, Sandeep Choubey, Mohammed Saleem","doi":"10.1101/2024.09.04.611260","DOIUrl":"https://doi.org/10.1101/2024.09.04.611260","url":null,"abstract":"Pathogenic bacteria evade host defense by rupturing the phagosomal compartment, enabling their escape into the host cytosol. The bacteria is known to establish direct physical contact with the host compartment prior to phagosome disruption. However, the impact of direct-physical contact on the phagosome remodeling and deformation remains elusive. To probe this, we first developed a method wherein we reconstitute a phagosome-like giant compartment encapsulating Mycobacterium smegmatis, a non-motile opportunistic bacterium. We discover that the direct contact between the bacteria and the encapsulating host membrane induces membrane bending, lipid wrapping, and local lipid phase separation at the contact site. The degree of phase separation is driven by the bacterial load leading to fluidization of the membrane, as evident from the decreased area stretch and bending modulus, making the host compartment more deformable. Surprisingly, for saturating bacterial load the fluid host membrane transforms into a scaffold-like rigid layer. We also find that the direct contact of the bacteria enhances the membranolytic potential of ESAT-6 thus contributing to its virulence. Together our findings provide mechanistic insights into the role of direct physical contact of the bacteria during phagosome disruption.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178216","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Membrane-assisted Aβ40 aggregation pathways 膜辅助 Aβ40 聚合途径

bioRxiv - Biophysics Pub Date : 2024-09-08 DOI: 10.1101/2024.09.05.611426

Fidha Nazreen Kunnath Muhammedkutty, Huan-Xiang Zhou

引用次数: 0

Grb2 Phosphorylation Antagonizes EGFR-driven Ras Activation by Interfering with Condensate Assembly Grb2 磷酸化通过干扰凝结物的组装来拮抗表皮生长因子受体驱动的 Ras 激活

bioRxiv - Biophysics Pub Date : 2024-09-07 DOI: 10.1101/2024.09.05.611544

Henry T Phan, Chun-Wei Lin, Brittany L. Stinger, Joseph B. DeGrandchamp, L.J. Nugent Lew, Serena J. Huang, Jay T Groves

{"title":"Grb2 Phosphorylation Antagonizes EGFR-driven Ras Activation by Interfering with Condensate Assembly","authors":"Henry T Phan, Chun-Wei Lin, Brittany L. Stinger, Joseph B. DeGrandchamp, L.J. Nugent Lew, Serena J. Huang, Jay T Groves","doi":"10.1101/2024.09.05.611544","DOIUrl":"https://doi.org/10.1101/2024.09.05.611544","url":null,"abstract":"Upon ligand binding, the kinase domain of EGFR phosphorylates multiple tyrosine residues on the receptor cytoplasmic tail through a trans-autophosphorylation process. Phosphotyrosine sites on activated receptors recruit Grb2, which further recruits SOS to initiate downstream signaling by activating Ras. Multivalent binding between SOS and Grb2, as well as direct Grb2:Grb2 interactions, contribute to formation of a protein condensate of activated EGFR. The condensed state of EGFR facilitates autoinhibition release in SOS and exerts regulatory control over signal propagation from activated EGFR to Ras. While kinase activity of EGFR is an essential driver of this signaling process, phosphorylation at residue Y160 on Grb2 blocks Grb2:Grb2 binding and can interfere with EGFR condensation. Here, using a reconstituted system, we examine how titrating kinase activity in the EGFR system can both promote and inhibit signal output to Ras. The results reveal how effects of tyrosine kinase inhibition can, under some circumstances, promote Ras activation by inhibiting negative feedback through Grb2 phosphorylation and disruption of a Grb2 SH2/SH3 dimer interface.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178254","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Modelling how lamellipodia-driven cells maintain persistent migration and interact with external barriers 模拟叶状薄片驱动的细胞如何保持持续迁移并与外部障碍相互作用

bioRxiv - Biophysics Pub Date : 2024-09-07 DOI: 10.1101/2024.09.06.611667

Shubhadeep Sadhukhan, Cristina Martinez-Torres, Samo Penic, Carsten Beta, Ales Iglic, Nir S. gov

{"title":"Modelling how lamellipodia-driven cells maintain persistent migration and interact with external barriers","authors":"Shubhadeep Sadhukhan, Cristina Martinez-Torres, Samo Penic, Carsten Beta, Ales Iglic, Nir S. gov","doi":"10.1101/2024.09.06.611667","DOIUrl":"https://doi.org/10.1101/2024.09.06.611667","url":null,"abstract":"Cell motility is fundamental to many biological processes, and cells exhibit a variety of migration patterns. Many motile cell types follow a universal law that connects their speed and persistency, a property that can originate from the intracellular transport of polarity cues due to the global actin retrograde flow. This mechanism was termed the ``Universal Coupling between cell Speed and Persistency\"(UCSP). Here we implemented a simplified version of the UCSP mechanism in a coarse-grained ``minimal-cell\" model, which is composed of a three-dimensional vesicle that contains curved active proteins. This model spontaneously forms a lamellipodia-like motile cell shape, which is however sensitive and can depolarize into a non-motile form due to random fluctuations or when interacting with external obstacles. The UCSP implementation introduces long-range inhibition, which stabilizes the motile phenotype. This allows our model to describe the robust polarity observed in cells and explain a large variety of cellular dynamics, such as the relation between cell speed and aspect ratio, cell-barrier scattering, and cellular oscillations in different types of geometric confinements.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-09-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142178255","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Nonequilibrium phases of a biomolecular condensate facilitated by enzyme activity 酶活性促进生物分子凝聚物的非平衡相

bioRxiv - Biophysics Pub Date : 2024-08-11 DOI: 10.1101/2024.08.11.607499

Sebastian T Coupe, Nikta Fakhri

{"title":"Nonequilibrium phases of a biomolecular condensate facilitated by enzyme activity","authors":"Sebastian T Coupe, Nikta Fakhri","doi":"10.1101/2024.08.11.607499","DOIUrl":"https://doi.org/10.1101/2024.08.11.607499","url":null,"abstract":"Biomolecular condensates represent a frontier in cellular organization, existing as dynamic materials driven out of equilibrium by active cellular processes. Here we explore active mechanisms of condensate regulation by examining the interplay between DEAD-box helicase activity and RNA base-pairing interactions within ribonucleoprotein condensates. We demonstrate how the ATP-dependent activity of DEAD-box helicases—a key class of enzymes in condensate regulation—acts as a nonequilibrium driver of condensate properties through the continuous remodeling of RNA interactions. By combining the LAF-1 DEAD-box helicase with a designer RNA hairpin concatemer, we unveil a complex landscape of dynamic behaviors, including time-dependent alterations in RNA partitioning, evolving condensate morphologies, and shifting condensate dynamics. Importantly, we reveal an antagonistic relationship between RNA secondary structure and helicase activity which promotes condensate homogeneity via a nonequilibrium steady state. By elucidating these nonequilibrium mechanisms, we gain a deeper understanding of cellular organization and expand the potential for active synthetic condensate systems.","PeriodicalId":501048,"journal":{"name":"bioRxiv - Biophysics","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-08-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141945053","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0