Biochimica et biophysica acta. Biomembranes最新文献_第6页

Novel flavonoid-magnesium complexes as inhibitors of plasma membrane calcium ATPase 新型类黄酮-镁复合物作为质膜钙atp酶抑制剂

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-08-05 DOI: 10.1016/j.bbamem.2025.184438

Mallku Q. Ontiveros , Mariel Marder , Rolando C. Rossi , Juan Pablo Rossi , Irene C. Mangialavori , Mariela S. Ferreira-Gomes

Plasma membrane calcium ATPases (PMCAs) are essential for regulating intracellular calcium (Ca²⁺) levels by extruding it from the cytosol. Improper regulation of these transporters is associated with numerous diseases, including neurological, cardiovascular, oncological, and metabolic problems, rendering them interesting targets for therapeutic intervention. However, there is a scarcity of specific tools to adjust PMCA activity. Flavonoids, a varied group of polyphenolic compounds with numerous biological effects, have been demonstrated to affect the function of several ATPases, including PMCAs.

In this study, we investigated the inhibitory mechanism of quercetin on the human PMCA4 isoform (hPMCA4). Using UV–visible spectroscopy and ATPase activity assay, we identified a high-affinity inhibition mediated by a quercetin‑magnesium (Mg²⁺) complex with a Ki of 49.7 ± 1.5 nM. Functional and phosphorylation studies at different pHs suggest that quercetin affects PMCA activity through two inhibitory mechanisms: a high-affinity one mediated by the quercetin-Mg²⁺ complex and a low-affinity one mediated by the free flavonoid.

Analysis of the structure-activity relationship revealed that hydroxyl groups at positions 3′, 4′, and 3 are critical for complex formation and inhibitory potency. Specifically, the 3′ and 4′ hydroxyls are required to form the PMCA inhibitory complex. These findings demonstrate a novel mechanism of PMCA activity modulation involving flavonoid-Mg²⁺ complexes, which emerge as selective molecular tools capable of regulating Ca²⁺ transport. This knowledge provides new insights into designing PMCA inhibitors and exploring therapeutic strategies targeting diseases linked to calcium signalling dysfunction.

质膜钙atp酶（PMCAs）是必不可少的调节细胞内钙（Ca2+）水平通过挤出它从细胞质溶胶。这些转运蛋白的不当调控与许多疾病有关，包括神经、心血管、肿瘤和代谢问题，使它们成为治疗干预的有趣靶点。然而，缺乏特定的工具来调整PMCA活动。黄酮类化合物是一类具有多种生物效应的多酚类化合物，已被证明可以影响几种atp酶的功能，包括PMCAs。在本研究中，我们研究了槲皮素对人PMCA4亚型（hPMCA4）的抑制机制。通过紫外可见光谱和atp酶活性测定，我们发现槲皮素-镁（Mg2+）配合物介导的高亲和力抑制作用，Ki为49.7±1.5 nM。不同ph值下的功能和磷酸化研究表明，槲皮素通过两种抑制机制影响PMCA活性：槲皮素- mg2 +复合物介导的高亲和力抑制机制和游离类黄酮介导的低亲和力抑制机制。构效关系分析表明，3′、4′和3位的羟基对复合物的形成和抑制作用至关重要。具体来说，3 ‘和4 ’羟基是形成PMCA抑制复合物所必需的。这些发现证明了PMCA活性调节的新机制，涉及黄酮类- mg2 +复合物，这是一种能够调节Ca2+运输的选择性分子工具。这些知识为设计PMCA抑制剂和探索针对钙信号功能障碍相关疾病的治疗策略提供了新的见解。

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引用次数: 0

Rescue of sirtuin inhibitor-dependent decrease in claudin-4 expression and paracellular barrier property in keratinocytes by epigallocatechin gallate 没食子儿茶素没食子酸酯对sirtuin抑制剂依赖性的角化细胞claudin-4表达降低和细胞旁屏障特性的拯救作用

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-08-06 DOI: 10.1016/j.bbamem.2025.184440

Maika Miwa , Miki Tanabe , Shunsuke Matsuda , Kento Yamakawa , Yuta Yoshino , Norihiro Tada , Akichika Itoh , Akira Ikari

The barrier function of granular layer in the skin is mainly sustained by claudin-1 (CLDN1) and CLDN4, tight junctional components. We recently found that the activity of sirtuin-2 (SIRT2), an anti-aging molecule, is decreased with aging in keratinocytes, leading to the attenuation of CLDN4 expression and paracellular barrier function. SIRT2 may be a novel target for enhancing skin barrier function in elderly people. In vitro SIRT2 activity assay showed that epigallocatechin gallate (EGCG) and green tea extract (GT) have a potent ability to activate SIRT2. Tenovin-1 (Ten-1), a sirtuin-1/2 inhibitor, decreased the SIRT2 activity in human keratinocyte-derived HaCaT cells, which was rescued by EGCG and GT. Ten-1 decreased the protein level of CLDN4, which was rescued by EGCG, whereas CLDN1 expression was changed by neither Ten-1 nor EGCG. Ten-1 decreased the tight junctional localization of CLDN4, transepithelial electrical resistance, and paracellular permeability to FD4, a fluorescence paracellular flux marker, which were rescued by EGCG. Ten-1 increased the acetylation level of CLDN4, which was inhibited by EGCG without affecting NAD⁺ content, a substrate for SIRT2. The protein levels of wild-type and K191A mutant were decreased by Ten-1, whereas that of K196A was not. Furthermore, Ten-1 increased the acetylation levels of WT and K191A mutant. We suggest that Ten-1 decreases CLDN4 expression mediated by the acetylation of K196 of CLDN4 and EGCG is useful to protect from aging-induced dysfunction of paracellular barrier in the keratinocytes.

皮肤颗粒层的屏障功能主要由紧密连接成分CLDN1 （CLDN1）和CLDN4维持。我们最近发现，抗衰老分子sirtuin-2 （SIRT2）的活性随着角质形成细胞的衰老而降低，导致CLDN4表达和细胞旁屏障功能的衰减。SIRT2可能是增强老年人皮肤屏障功能的新靶点。体外SIRT2活性分析表明，表没食子儿茶素没食子酸酯（EGCG）和绿茶提取物（GT）具有有效的SIRT2激活能力。Tenovin-1 （Ten-1）是一种sirtuin-1/2抑制剂，可降低EGCG和GT拯救的人角化细胞来源的HaCaT细胞中SIRT2的活性。Ten-1可降低EGCG拯救的CLDN4的蛋白水平，而CLDN1的表达不受Ten-1和EGCG的影响。10 -1降低了CLDN4的紧密连接定位、经上皮电阻和细胞旁对FD4（一种荧光细胞旁通量标记物）的通透性，这些被EGCG拯救。10 -1增加了CLDN4的乙酰化水平，EGCG抑制了CLDN4的乙酰化，但不影响SIRT2的底物NAD+含量。野生型和K191A突变体的蛋白水平下降了10 -1，而K196A突变体的蛋白水平没有下降。此外，10 -1增加了WT和K191A突变体的乙酰化水平。我们认为，10 -1降低CLDN4和EGCG的K196乙酰化介导的CLDN4表达，有助于保护角质形成细胞免受衰老诱导的细胞旁屏障功能障碍。

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引用次数: 0

Two phase coexistence in ternary mixtures of saturated and polyunsaturated phosphatidylcholines with cholesterol. 饱和和多不饱和磷脂酰胆碱与胆固醇三元混合物的两相共存。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-08-06 DOI: 10.1016/j.bbamem.2025.184436

James H Davis

Combining ²H and ¹⁴N nuclear magnetic resonance measurements on ternary lipid/cholesterol mixtures it is possible to quantitate the phase separation into coexisting ℓ_o and ℓ_d domains using a single sample whose composition is within the two phase region. Mixtures of each of the unsaturated phospholipids dioleoyl phosphatidylcholine, dilinoleoyl phosphatidylcholine and dilinolenoyl phosphatidylcholine with the saturated dipalmitoyl phosphatidylcholine and cholesterol all exhibit ℓ_o and ℓ_d phase coexistence over a substantial range of compositions and temperatures. A higher degree of unsaturation broadens the temperature range of two phase coexistence and results in the ℓ_d phase domains actually being significantly more 'fluid' at lower temperatures than they are at higher temperatures.

结合对三元脂/胆固醇混合物的2H和14N核磁共振测量，可以使用一个组成在两相区域内的单一样品来量化相分离到共存的_o和_d域。每一种不饱和磷脂二酰磷脂酰胆碱、二酰磷脂酰胆碱和二酰磷脂酰胆碱与饱和二棕榈酰磷脂酰胆碱和胆固醇的混合物在相当大的组成和温度范围内均表现出0和d相共存。较高的不饱和程度扩大了两相共存的温度范围，并导致在较低温度下的d相域实际上比在较高温度下更具有“流动性”。

引用次数: 0

The phospholipolytically active neurotoxin Vipoxin induces changes of the mechanical properties of breast epithelial cells 磷脂多活性神经毒素Vipoxin诱导乳腺上皮细胞力学特性的改变。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-07-09 DOI: 10.1016/j.bbamem.2025.184434

Will Linthicum , Qi Wen , Nancy A. Burnham , Svetla D. Petrova , Konstantin Balashev

The investigation of how drugs or toxins alter cell mechanics is gaining significant traction in biomedical science, driven by the dual objectives of elucidating their mechanisms of action and enhancing drug screening processes. In this study, Atomic Force Microscopy (AFM), a prominent experimental technique in recent years, was employed to examine and analyze the mechanical responses of cells exposed to the neurotoxin Vipoxin. This method enables the precise measurement of key mechanical parameters such as cell stiffness and viscoelasticity before and after toxin introduction in the cell culture. It was demonstrated that the cells' stiffness and viscosity decreased with increasing Vipoxin concentration. Additionally, Total Internal Reflection Fluorescence Microscopy (TIRFM) was utilized to monitor morphological changes in the cells over time. These morphological changes were quantitatively analyzed using fractal analysis of the acquired images. The observed changes in cell shapes implied the reorganization of the cell cytoskeleton, thus providing insight into a comprehensive understanding of cell mechanics under the influence of Vipoxin.

在阐明其作用机制和加强药物筛选过程的双重目标的驱动下，药物或毒素如何改变细胞力学的研究在生物医学科学中获得了显著的吸引力。在本研究中，原子力显微镜（AFM）是近年来一项重要的实验技术，用于检测和分析细胞暴露于神经毒素Vipoxin的力学反应。该方法能够在细胞培养中引入毒素前后精确测量细胞刚度和粘弹性等关键力学参数。结果表明，随着Vipoxin浓度的增加，细胞的刚度和黏度降低。此外，利用全内反射荧光显微镜（TIRFM）监测细胞随时间的形态学变化。这些形态变化是定量分析使用分形分析获得的图像。观察到的细胞形状的变化暗示了细胞骨架的重组，从而为全面了解Vipoxin影响下的细胞力学提供了见解。

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引用次数: 0

Development of amyloid-cationic peptides with antimicrobial activities: Relation to their membranotropic activities. 具有抗菌活性的淀粉样阳离子肽的开发：与其膜性活性的关系。

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-08-07 DOI: 10.1016/j.bbamem.2025.184439

Marta Balestra, Lilia Leghmizi, Thierry Drujon, Loïc Portier, Cillian Byrne, Fabienne Burlina, Sylvie Noinville

We describe here a new class of antimicrobial peptides (named Amy-Cat) comprised of a short amyloid domain and a cationic domain, as a primary amphipathic structure. The nona-arginine sequence was chosen as the cationic motif, while the sequence and size of the amyloid domain was modulated. The Amy-Cat peptides were found to be bactericidal against gram-negative and gram-positive standard bacterial strains with minimum inhibitory concentrations ranging from 3 to 24 μM, and being well-below their hemolytic concentrations. Their membranotropic activities were investigated as a function of the amyloid sequence and compared to those of the nona-arginine peptide. Calcein dye leakage on lipid mimic models for bacterial and eukaryotic membranes was carried out. In addition, the effect of the amyloid moiety on the membrane binding and on the conformational change were investigated at the buffer/supported lipid bilayer interface using ATR-FTIR spectroscopy. The overall findings suggest optimum routes to balancing the hydrophobicity of the amyloid sequence over the fixed cationic sequence allowing selective disruption of the bacterial membranes without eliciting hemolysis. Amy-Cat peptides appear to be very promising candidates for the development of new antimicrobial agents.

我们在这里描述了一类新的抗菌肽（命名为Amy-Cat），它由一个短淀粉样结构域和一个阳离子结构域组成，是一个主要的两亲结构。选择非精氨酸序列作为阳离子基序，同时调节淀粉样结构域的序列和大小。研究发现，ami - cat肽对革兰氏阴性和革兰氏阳性标准菌株具有杀菌作用，其最低抑菌浓度为3 ~ 24 μM，远低于其溶血浓度。它们的膜性活性作为淀粉样蛋白序列的功能进行了研究，并与nona-精氨酸肽进行了比较。研究了钙黄蛋白在细菌和真核生物膜脂质模拟模型上的渗漏。此外，利用ATR-FTIR光谱研究了淀粉样蛋白片段对缓冲/支撑脂质双层界面上膜结合和构象变化的影响。总的研究结果表明，平衡淀粉样蛋白序列在固定阳离子序列上的疏水性的最佳途径，允许在不引起溶血的情况下选择性地破坏细菌膜。Amy-Cat肽似乎是开发新型抗菌药物的非常有前途的候选者。

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引用次数: 0

Assessing the anticancer potential of spider venom peptide Latarcin Ltc2a against triple negative breast cancer 评估蜘蛛毒液肽Latarcin Ltc2a对三阴性乳腺癌的抗癌潜力

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-08-20 DOI: 10.1016/j.bbamem.2025.184442

Prasanjeet Kaur , Srabaita Roy , Shilpi Minocha , Archana Chugh

Cancer remains one of the most formidable challenges to human health, necessitating constant exploration of innovative therapeutic strategies. Among the myriad potential candidates, peptides from venom have emerged as potent sources of bioactive molecules possessing diverse pharmacological properties. In this study, we repurposed a spider venom-derived antimicrobial peptide, Ltc2a, into a selective anticancer agent, bridging microbial defense with cancer therapeutics. Our findings reveal that Ltc2a exhibits selective cytotoxicity towards cancer cells compared to normal cells at just 2 μM of the peptide concentration. Ltc2a induced rapid cytotoxicity within 1 h in breast cancer cells and it was accompanied by membrane disruption as shown by propidium iodide (PI) positive staining and visible damage to cancer cell membranes under field emission scanning electron microscopy (FESEM). In vivo studies using a zebrafish model indicated favorable uptake and a lack of acute toxicity, depicting 80 % survival rate up to 4 μM of tested peptide concentration. Interestingly, the truncated variants of Ltc2a retained their alpha helical structure and demonstrated preferential uptake in MDA-MB-231 cells over HEK293T cells. These findings highlight the therapeutic potential of Ltc2a as selective anticancer peptide with minimal toxicity, paving the way for further preclinical development.

癌症仍然是人类健康面临的最严峻挑战之一，需要不断探索创新的治疗策略。在众多潜在的候选者中，来自毒液的肽已成为具有多种药理特性的生物活性分子的有力来源。在这项研究中，我们将蜘蛛毒液衍生的抗菌肽Ltc2a改造成一种选择性抗癌剂，将微生物防御与癌症治疗联系起来。我们的研究结果表明，与正常细胞相比，Ltc2a在2 μM的肽浓度下对癌细胞表现出选择性的细胞毒性。Ltc2a在1 h内对乳腺癌细胞产生快速的细胞毒性，并伴有膜破坏，如碘化丙啶（PI）阳性染色和场发射扫描电镜（FESEM）下可见的癌细胞膜损伤。使用斑马鱼模型进行的体内研究表明，在4 μM的测试肽浓度下，其具有良好的吸收性和无急性毒性，存活率为80%。有趣的是，Ltc2a的截断变体保留了其α螺旋结构，并且在MDA-MB-231细胞中表现出比HEK293T细胞更优先的摄取。这些发现突出了Ltc2a作为毒性最小的选择性抗癌肽的治疗潜力，为进一步的临床前开发铺平了道路。

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引用次数: 0

Investigation of biophysical properties of ion channels with nonlinear methods 用非线性方法研究离子通道的生物物理性质

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-07-30 DOI: 10.1016/j.bbamem.2025.184437

Mahmut Akilli , Fatma Söğüt , Ülkü Çömelekoğlu , Handan Tuncel

The aim of this study is to show how nonlinear methods can be used to investigate the biophysical properties of ion channels. For this purpose, the membrane ion current signals of the EAG1 potassium channel of MCF-7 cells and of the TRP channel of ARPE-19 cells were used. Entropy measurements and maximum Lyapunov exponent were chosen as nonlinear methods. The vital state functional of the ion channels in the membrane was monitored using the entropy parameter. The behavioural or functional sensitivity of ion channels was quantified by the maximum Lyapunov exponent. It is known that the entropy of a system increases as it moves towards equilibrium. In this context, during the electrical activity of a living cell, the entropy of the cell reaches its maximum when the membrane ion fluxes reach the equilibrium, that is, when the value of the ion fluxes approaches zero. Therefore, the accuracy of the results obtained in this study was calibrated by reference to this general assumption. The results show functional differences between the MCF-7 EAG1 potassium channel and the ARPE-19 TRP channel. This method has potential applications in analysing cell behaviour or studying ion channel biophysical properties. It can also be used to observe differences in the behaviour of normal and cancerous cells of the same type, or to measure the effects of drugs on the cell.

本研究的目的是展示如何使用非线性方法来研究离子通道的生物物理特性。为此，我们使用了MCF-7细胞EAG1钾通道和ARPE-19细胞TRP通道的膜离子电流信号。选择熵测量和最大李雅普诺夫指数作为非线性方法。利用熵参数对膜内离子通道的动态功能进行了监测。用最大李亚普诺夫指数量化离子通道的行为或功能敏感性。众所周知，当系统趋向平衡时，它的熵增加。在这种情况下，在活细胞的电活动过程中，当膜离子通量达到平衡时，即当离子通量的值接近于零时，细胞的熵达到最大值。因此，本研究所得结果的准确性参照这一一般假设进行校准。结果显示MCF-7 EAG1钾通道和ARPE-19 TRP通道在功能上存在差异。该方法在分析细胞行为或研究离子通道生物物理性质方面具有潜在的应用前景。它还可以用来观察同种正常细胞和癌细胞的行为差异，或测量药物对细胞的影响。

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引用次数: 0

Accurate antibiotic accumulation in Enterobacteriaceae isolates expressing efflux pumps 表达外排泵的肠杆菌科分离物中抗生素的准确积累

IF 2.5 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-10-01 Epub Date: 2025-08-08 DOI: 10.1016/j.bbamem.2025.184441

Jinane Tabcheh , Julia Vergalli , Jean-Marie Pagès , Jean Michel Brunel

In Enterobacteriaceae, antibiotic susceptibility is frequently influenced by mechanisms such as membrane modifications, target site mutations, and enzymatic resistance barriers. Recently, there has been a notable rise in Klebsiella pneumoniae, Escherichia coli, and Enterobacter cloacae isolates exhibiting antibiotic resistance in hospital settings. Of particular concern, some resistant isolates employ membrane-associated resistance mechanisms that significantly lower intracellular antibiotic concentrations, reducing them below the threshold required for therapeutic efficacy. Advancements in methods for quantifying drug accumulation within bacterial cells have provided critical insights into these resistance mechanisms. A key step in these studies relies on cell lysis to release intracellular contents including antibacterial molecules for precise quantification. However, current lysis methods are often time-consuming, underscoring the need for a robust, efficient approach to accurately measure intracellular antibiotic concentrations in isolates exhibiting various levels of efflux pump activity. In this study, we developed a rapid and reliable lysis protocol that minimizes the risk of drug alteration while enabling precise and reproducible measurement of intracellular antibiotic concentrations allowing an evidence-based study of efflux in resistant clinical strains of Enterobacteriaceae. This approach holds significant promise for enhancing our understanding of membrane-associated resistance mechanisms and for informing the optimization of treatment strategies.

在肠杆菌科中，抗生素敏感性经常受到膜修饰、靶点突变和酶抗性屏障等机制的影响。最近，在医院环境中肺炎克雷伯菌、大肠杆菌和阴沟肠杆菌分离株表现出抗生素耐药性的病例显著增加。特别值得关注的是，一些耐药菌株采用膜相关耐药机制，显著降低细胞内抗生素浓度，使其低于治疗效果所需的阈值。定量细菌细胞内药物积累方法的进步为这些耐药机制提供了重要的见解。这些研究的关键步骤依赖于细胞裂解释放细胞内内容物，包括抗菌分子，以进行精确定量。然而，目前的裂解方法往往是耗时的，强调需要一个强大的，有效的方法来准确测量细胞内抗生素浓度的分离表现出不同水平的外排泵活性。在这项研究中，我们开发了一种快速可靠的裂解方案，最大限度地降低了药物改变的风险，同时能够精确和可重复地测量细胞内抗生素浓度，从而对肠杆菌科耐药临床菌株的外排进行循证研究。这种方法对增强我们对膜相关耐药机制的理解和优化治疗策略具有重要的意义。

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引用次数: 0

Effect of membrane tension on (−)-epigallocatechin gallate-induced burst of single giant unilamellar vesicles 膜张力对(-)-表没食子儿茶素没食子酸盐诱导的单层大囊泡破裂的影响。

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-06-01 Epub Date: 2025-05-26 DOI: 10.1016/j.bbamem.2025.184427

Yukihiro Tamba , Naoya Sugita , Mika Terada , Masahito Yamazaki

(−)-Epigallocatechin gallate (EGCg), a tea catechin, exhibits antimicrobial activity. EGCg induces burst of giant unilamellar vesicles (GUVs), resulting in leakage of their internal contents. Here, we examined the effect of membrane tension on the EGCg-induced burst of dioleoylphosphatidylcholine (DOPC)-GUVs. The observation of the EGCg-induced burst of GUVs without membrane tension indicated that first a micropore was formed in the membrane and then its radius rapidly increased with time within ~10 ms without a change in the GUV diameter, and then the GUV diameter decreased to a lipid membrane aggregate. Next, we examined the effect of membrane tension on the EGCg-induced fractional area change (δ) of GUVs and its burst. During the interaction of EGCg with a GUV with low tension (≤ 0.5 mN/m), the δ initially increased slightly and then decreased to negative values, concomitant with the appearance of invaginated structures such as dense particles and narrow tubes in the GUV membrane and lumen, resulting in pore formation and subsequent GUV burst. By contrast, at higher tension, δ increased with time. The fraction of burst GUV after 5 min of interaction, P_burst (5 min), decreased with increasing tension up to 3.0 mN/m, indicating that membrane tension suppressed the burst. The P_burst (5 min) increased with increasing the fraction of GUVs in which invaginated structures appeared within 5 min of interaction, suggesting that the formation of invaginated structures may cause the initial EGCg-induced pore formation. We have proposed a mechanism of the tension effect on the EGCg-induced pore formation.

（-）-表没食子儿茶素没食子酸酯（EGCg），茶儿茶素，具有抗菌活性。EGCg诱导巨大单层囊泡（GUVs）破裂，导致其内部内容物泄漏。在这里，我们检测了膜张力对卵磷脂诱导的二油基磷脂酰胆碱(DOPC)- guv爆发的影响。在无膜张力的情况下，egcg诱导的GUV破裂观察表明，在~10 ms内，膜上先形成微孔，微孔半径随时间迅速增大，但GUV直径没有变化，随后GUV直径减小为脂质膜聚集体。接下来，我们研究了膜张力对egcg诱导的guv分数面积变化（δ）及其破裂的影响。在EGCg与低张力（≤0.5 mN/m）的GUV相互作用过程中，δ先小幅升高，后降至负值，GUV膜和管腔内出现致密颗粒、窄管等内翻结构，形成孔洞，导致GUV破裂。在较高的张力下，δ随时间增加而增加。当膜张力达到3.0 mN/m时，作用5 min后破裂GUV的比例Pburst（5 min）随着膜张力的增加而降低，表明膜张力抑制了破裂。在相互作用的5 min内，Pburst（5 min）随着出现内陷结构的guv比例的增加而增加，表明内陷结构的形成可能是egcg诱导的初始孔隙形成的原因。我们提出了一种张力作用于卵磷脂诱导的孔隙形成的机制。

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引用次数: 0

Acidic bicelles are a suitable membrane mimic for structural studies of the Lassa virus fusion domain 酸性单胞体是拉沙病毒融合结构域结构研究的合适膜模拟物

IF 2.8 3区生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY

Biochimica et biophysica acta. Biomembranes

Pub Date : 2025-06-01 Epub Date: 2025-06-05 DOI: 10.1016/j.bbamem.2025.184428

Hallie N. Pennington, Jinwoo Lee

Lassa virus (LASV) is the most prevalent arenavirus afflicting humans and has high pandemic potential. The genetic material of LASV is delivered into the host cell via membrane fusion initiated by the LASV fusion domain (FD). However, the molecular details of the LASV FD, particularly its structure after association with the host cell, remain poorly characterized. This can be attributed to a lack of a viable membrane mimic to effectively stabilize the LASV FD for structural studies. Here, we demonstrate that the structure of the LASV FD widely varies based on the class of membrane mimic. In particular, through CD spectroscopy, we found that the LASV FD required a charged membrane mimic, such as zwitterionic or anionic detergent micelles, to adopt a helical conformation at low pH, but has the highest helical content in the presence of anionic lipids, particularly the detergent micelle LMPG and acidic bicelles. Moreover, we reveal that the LASV FD was well resolved on NMR spectra in CHAPS, DPC, LDAO, LMPG, and acidic bicelles, where LMPG and acidic bicelles had the sharpest peak resolution, but more defined peaks were noted in acidic bicelles over LMPG. In conclusion, our findings indicate that acidic bicelles are the optimal membrane mimic for the stabilization of the LASV FD such that structural studies can be conducted.

拉沙病毒（LASV）是影响人类最普遍的沙粒病毒，具有很高的大流行潜力。LASV的遗传物质通过由LASV融合域（FD）启动的膜融合传递到宿主细胞中。然而，LASV FD的分子细节，特别是其与宿主细胞结合后的结构，仍然缺乏表征。这可归因于缺乏可行的膜模拟物来有效地稳定LASV FD用于结构研究。在这里，我们证明了LASV FD的结构根据膜模拟物的类别有很大的不同。特别是，通过CD光谱，我们发现LASV FD需要带电的膜模拟物，如两性离子或阴离子洗涤剂胶束，在低pH下采用螺旋构象，但在阴离子脂质存在时，特别是洗涤剂胶束LMPG和酸性单束存在时，螺旋含量最高。此外，我们发现LASV FD在CHAPS、DPC、LDAO、LMPG和酸性小柱的核磁共振光谱上被很好地分辨出来，其中LMPG和酸性小柱的峰分辨率最高，而LMPG的酸性小柱的峰更清晰。总之，我们的研究结果表明，酸性双胞体是稳定LASV FD的最佳膜模拟物，因此可以进行结构研究。

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引用次数: 0