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Identification of a new DC-SIGN binding pentamannoside epitope within the complex structure of Candida albicans mannan 甘露白色念珠菌复杂结构中一个新的DC-SIGN结合五甘露糖苷表位的鉴定
Q1 Immunology and Microbiology Pub Date : 2023-07-22 DOI: 10.1016/j.tcsw.2023.100109
Vadim B. Krylov , Marcos Gómez-Redondo , Arsenii S. Solovev , Dmitry V. Yashunsky , Alistair J.P. Brown , Mark H.T. Stappers , Neil A.R. Gow , Ana Ardá , Jesús Jiménez-Barbero , Nikolay E. Nifantiev

The dendritic cell-specific intercellular adhesion molecule-3-grabbing non-integrin (DC-SIGN) is an innate immune C-type lectin receptor that recognizes carbohydrate-based pathogen associated with molecular patterns of various bacteria, fungi, viruses and protozoa. Although a range of highly mannosylated glycoproteins have been shown to induce signaling via DC-SIGN, precise structure of the recognized oligosaccharide epitope is still unclear. Using the array of oligosaccharides related to selected fragments of main fungal antigenic polysaccharides we revealed a highly specific pentamannoside ligand of DC-SIGN, consisting of α-(1 → 2)-linked mannose chains with one inner α-(1 → 3)-linked unit. This structural motif is present in Candida albicans cell wall mannan and corresponds to its antigenic factors 4 and 13b. This epitope is not ubiquitous in other yeast species and may account for the species-specific nature of fungal recognition via DC-SIGN. The discovered highly specific oligosaccharide ligands of DC-SIGN are tractable tools for interdisciplinary investigations of mechanisms of fungal innate immunity and anti-Candida defense. Ligand- and receptor-based NMR data demonstrated the pentasaccharide-to-DC-SIGN interaction in solution and enabled the deciphering of the interaction topology.

树突状细胞特异性细胞间粘附分子-3-抓非整联素(DC-SIGN)是一种先天免疫c型凝集素受体,可识别与各种细菌、真菌、病毒和原生动物分子模式相关的碳水化合物为基础的病原体。尽管一系列高甘露糖基化糖蛋白已被证明可通过DC-SIGN诱导信号传导,但被识别的寡糖表位的精确结构仍不清楚。利用与主要真菌抗原性多糖片段相关的低聚糖阵列,我们发现了DC-SIGN的高度特异性五氨基糖苷配体,由α-(1→2)连接的甘露糖链和一个内部α-(1→3)连接的单元组成。该结构基序存在于白色念珠菌细胞壁甘露聚糖中,与其抗原因子4和13b相对应。这个表位在其他酵母物种中并不普遍存在,这可能解释了真菌通过DC-SIGN识别的物种特异性。所发现的DC-SIGN高特异性低聚糖配体为真菌先天免疫和抗念珠菌防御机制的跨学科研究提供了方便的工具。基于配体和受体的核磁共振数据证明了溶液中五糖与dc - sign的相互作用,并能够破译相互作用的拓扑结构。
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引用次数: 1
Binding of micro-nutrients to the cell wall of the fungus Schizophyllum commune 微量营养物质与真菌裂褶菌细胞壁的结合
Q1 Immunology and Microbiology Pub Date : 2023-06-17 DOI: 10.1016/j.tcsw.2023.100108
Fleur E.L. Kleijburg , Adil A. Safeer , Marc Baldus , Han A.B. Wösten

The cell wall fulfils several functions in the biology of fungi. For instance, it provides mechanical strength, interacts with the (a)biotic environment, and acts as a molecular sieve. Recently, it was shown that proteins and β-glucans in the cell wall of Schizophyllum commune bind Cu2+. We here show that the cell wall of this mushroom forming fungus also binds other (micro-)nutrients. Ca2+, Mg2+, Mn2+, NO3, PO43-, and SO42- bound at levels > 1 mg per gram dry weight cell wall, while binding of BO3-, Cu2+, Zn2+ and MoO42- was lower. Sorption of Ca2+, Mn2+, Zn2+ and PO43- was promoted at alkaline pH. These compounds as well as BO33-, Cu2+, Mg2+, NO3, and SO42- that had bound at pH 4, 6, or 8 could be released from the cell wall at pH 4 with a maximum efficiency of 46–93 %. Solid-state NMR spectroscopy showed that the metals had the same binding sites as Cu2+ when a low concentration of this ion is used. Moreover, data indicate that anions bind to the cell wall as well as to the metal ions. Together, it is shown that the cell wall of S. commune binds various (micro-)nutrients and that this binding is higher than the uptake by hyphae. The binding to the cell wall may be used as a storage mechanism or may reduce availability of these molecules to competitors or prevent toxic influx in the cytoplasm.

细胞壁在真菌生物学中具有多种功能。例如,它提供机械强度,与(a)生物环境相互作用,并充当分子筛。最近,人们发现裂褶菌细胞壁中的蛋白质和β-葡聚糖与Cu2+结合。我们在这里表明,这种蘑菇形成真菌的细胞壁也结合了其他(微量)营养物质。Ca2+、Mg2+、Mn2+、NO3-、PO43-和SO42-结合水平>;1mg/克干重细胞壁,而BO3-、Cu2+、Zn2+和MoO42-的结合较低。在碱性pH下,Ca2+、Mn2+、Zn2+和PO43-的吸附得到促进。这些化合物以及在pH 4、6或8时结合的BO33-、Cu2+、Mg2+、NO3-和SO42-在pH 4时可以从细胞壁释放,最大效率为46–93%。固态NMR光谱显示,当使用低浓度的Cu2+时,金属具有与Cu2+相同的结合位点。此外,数据表明阴离子与细胞壁结合,也与金属离子结合。总之,研究表明,S.community的细胞壁结合了各种(微量)营养物质,并且这种结合高于菌丝的吸收。与细胞壁的结合可以用作储存机制,或者可以减少这些分子对竞争对手的可用性,或者防止有毒物质流入细胞质。
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引用次数: 1
Strain and temperature dependent aggregation of Candida auris is attenuated by inhibition of surface amyloid proteins 菌株和温度依赖的耳念珠菌聚集是通过抑制表面淀粉样蛋白减弱
Q1 Immunology and Microbiology Pub Date : 2023-05-09 DOI: 10.1101/2023.05.09.540062
Dhara Malavia-Jones, Rhys A. Farrer, M. Stappers, Matt B. Edmondson, A. Borman, Elizabeth M. Johnson, P. Lipke, N. Gow
Candida auris is a multi-drug resistant human fungal pathogen that has become a global threat to human health due to its drug resistant phenotype, persistence in the hospital environment and propensity for patient to patient spread. Isolates display variable aggregation that may affect the relative virulence of strains. Therefore, dissection of this phenotype has gained substantial interest in recent years. We studied eight clinical isolates from four different clades (I-IV); four of which had a strongly aggregating phenotype and four of which did not. Genome analysis identified polymorphisms associated with loss of cell surface proteins were enriched in weakly-aggregating strains. Additionally, we identified down-regulation of chitin synthase and chitinase genes involved in the synthesis and dissolution of the chitinous septum. Characterisation of the cells revealed no ultrastructural defects in cytokinesis or cell separation in aggregating isolates. Strongly and weakly aggregating strains did not differ in net surface charge or in cell surface hydrophobicity. The capacity for aggregation and for adhesion to polystyrene microspheres were also not correlated. However, aggregation and extracellular matrix formation were all increased at higher growth temperatures, and treatment with the amyloid protein inhibitor Thioflavin-T markedly attenuated aggregation. Genome analysis further indicated strain specific differences in the genome content of GPI-anchored proteins including those encoding genes with the potential to form amyloid proteins. Collectively our data suggests that aggregation is a complex strain and temperature dependent phenomenon that may be linked in part to the ability to form extracellular matrix and cell surface amyloids. HIGHLIGHTS The amyloid inhibitor Thioflavin-T inhibited C. auris aggregation. Aggregating isolates do not exhibit any defects in cell separation. Genomic differences were identified between strongly aggregating and weakly-aggregating strains of C. auris. Aggregation did not correlate with surface charge or hydrophobicity of yeast cells.
耳念珠菌是一种耐多种药物的人类真菌病原体,由于其耐药性表型、在医院环境中的持久性和患者间传播的倾向,已成为对人类健康的全球威胁。分离株表现出可变的聚集性,这可能影响菌株的相对毒力。因此,近年来对这种表型的解剖引起了人们的极大兴趣。我们研究了来自四个不同分支(I-IV)的八个临床分离株;其中4个具有强聚集表型,4个没有。基因组分析发现,与细胞表面蛋白损失相关的多态性在弱聚集菌株中富集。此外,我们还鉴定了参与几丁质隔膜合成和溶解的几丁质合成酶和几丁质酶基因的下调。细胞的特征显示在胞质分裂或聚集分离物中没有细胞分离的超微结构缺陷。强和弱聚集菌株在净表面电荷或细胞表面疏水性方面没有差异。聚集能力和与聚苯乙烯微球的粘附能力也没有相关性。然而,在较高的生长温度下,聚集和细胞外基质的形成都增加了,用淀粉样蛋白抑制剂硫黄素-T处理显著减弱了聚集。基因组分析进一步表明,GPI锚定蛋白的基因组含量存在菌株特异性差异,包括编码有可能形成淀粉样蛋白的基因的差异。总的来说,我们的数据表明,聚集是一种复杂的菌株和温度依赖性现象,可能部分与形成细胞外基质和细胞表面淀粉样蛋白的能力有关。亮点淀粉样蛋白抑制剂硫黄素-T抑制金黄色葡萄球菌的聚集。聚集的分离物在细胞分离中没有表现出任何缺陷。在强聚集菌株和弱聚集菌株之间鉴定了基因组差异。聚集性与酵母细胞的表面电荷或疏水性无关。
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引用次数: 3
Erratum to “The nature of the fungal cargo induces significantly different temporal programmes of macrophage phagocytosis” [Cell Surf. 8 (2022) 100082] 对“真菌货物的性质诱导巨噬细胞吞噬的显著不同的时间程序”的勘误[Cell Surf. 8 (2022) 100082]
Q1 Immunology and Microbiology Pub Date : 2023-04-28 DOI: 10.1016/j.tcsw.2023.100106
María Fernanda Alonso , Judith M. Bain , Fiona M. Rudkin , Lars P. Erwig , Alistair J.P. Brown , Neil A.R. Gow
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引用次数: 0
Diffusion in intact secondary cell wall models of plants at different equilibrium moisture content 不同平衡含水量下植物完整次生细胞壁模型的扩散
Q1 Immunology and Microbiology Pub Date : 2023-03-25 DOI: 10.1016/j.tcsw.2023.100105
Daipayan Sarkar , Lintao Bu , Joseph E. Jakes , Jacob K. Zieba , Isaiah D. Kaufman , Michael F. Crowley , Peter N. Ciesielski , Josh V. Vermaas

Secondary plant cell walls are composed of carbohydrate and lignin polymers, and collectively represent a significant renewable resource. Leveraging these resources depends in part on a mechanistic understanding for diffusive processes within plant cell walls. Common wood protection treatments and biomass conversion processes to create biorefinery feedstocks feature ion or solvent diffusion within the cell wall. X-ray fluorescence microscopy experiments have determined that ionic diffusion rates are dependent on cell wall hydration as well as the ionic species through non-linear relationships. In this work, we use classical molecular dynamics simulations to map the diffusion behavior of different plant cell wall components (cellulose, hemicellulose, lignin), ions (Na+, K+, Cu2+, Cl) and water within a model for an intact plant cell wall at various hydration states (3–30 wt% water). From these simulations, we analyze the contacts between different plant cell wall components with each other and their interaction with the ions. Generally, diffusion increases with increasing hydration, with lignin and hemicellulose components increasing diffusion by an order of magnitude over the tested hydration range. Ion diffusion depends on charge. Positively charged cations preferentially interact with hemicellulose components, which include negatively charged carboxylates. As a result, positive ions diffuse more slowly than negatively charged ions. Measured diffusion coefficients are largely observed to best fit piecewise linear trends, with an inflection point between 10 and 15% hydration. These observations shed light onto the molecular mechanisms for diffusive processes within secondary plant cell walls at atomic resolution.

次生植物细胞壁由碳水化合物和木质素聚合物组成,共同代表了一种重要的可再生资源。利用这些资源在一定程度上取决于对植物细胞壁内扩散过程的机械理解。常见的木材保护处理和生物质转化过程以产生生物精炼原料,其特征是离子或溶剂在细胞壁内扩散。X射线荧光显微镜实验已经确定,离子扩散速率通过非线性关系取决于细胞壁水合作用以及离子种类。在这项工作中,我们使用经典的分子动力学模拟来绘制不同植物细胞壁成分(纤维素、半纤维素、木质素)、离子(Na+、K+、Cu2+、Cl−)和水在不同水合状态(3–30 wt%水)下完整植物细胞壁模型中的扩散行为。通过这些模拟,我们分析了不同植物细胞壁成分之间的相互接触及其与离子的相互作用。通常,扩散随着水合作用的增加而增加,木质素和半纤维素组分在测试的水合作用范围内扩散增加了一个数量级。离子的扩散取决于电荷。带正电荷的阳离子优先与半纤维素成分相互作用,半纤维素成分包括带负电荷的羧酸盐。因此,正离子比带负电的离子扩散得更慢。测量的扩散系数在很大程度上被观察到最适合分段线性趋势,拐点在10%和15%之间。这些观察结果揭示了在原子分辨率下次生植物细胞壁内扩散过程的分子机制。
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引用次数: 2
In vivo measurement of the Young’s modulus of the cell wall of single root hairs 单根毛细胞壁杨氏模量的体内测定
Q1 Immunology and Microbiology Pub Date : 2023-03-01 DOI: 10.1016/j.tcsw.2023.100104
David Pereira , Thomas Alline , Sébastjen Schoenaers , Atef Asnacios

Root hairs are cells from the root epidermis that grow as long tubular bulges perpendicular to the root. They can grow in a variety of mechanical or chemical environments. Their mechanical properties are mainly due to their stiff cell wall which also constitutes a physical barrier between the cell and its environment. Thus, it is essential to be able to quantify the cell wall mechanical properties and their adaptation to environmental cues. Here, we present a technique we developed to measure the Young’s (elastic) modulus of the root hair cell wall. In essence, using custom-made glass microplates as cantilevers of calibrated stiffness, we are able to measure the force necessary to bend a single living root hair. From these experiments one can determine the stiffness and Young’s modulus of the root hair cell wall.

根毛是来自根表皮的细胞,生长为垂直于根的长管状凸起。它们可以在各种机械或化学环境中生长。它们的机械性能主要是由于它们坚硬的细胞壁,这也构成了细胞与其环境之间的物理屏障。因此,能够量化细胞壁的机械特性及其对环境线索的适应是至关重要的。在这里,我们提出了一种我们开发的技术来测量根毛细胞壁的杨氏(弹性)模量。本质上,使用定制的玻璃微板作为校准刚度的悬臂,我们能够测量弯曲单个活根毛所需的力。从这些实验中可以确定根毛细胞壁的刚度和杨氏模量。
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引用次数: 1
Observing cellulose synthases at emerging secondary thickenings in developing xylem vessels of the plant root using airyscan confocal microscopy 利用空气扫描共聚焦显微镜观察植物根部木质部导管中出现的次生增厚中的纤维素合酶
Q1 Immunology and Microbiology Pub Date : 2023-02-28 DOI: 10.1016/j.tcsw.2023.100103
Raymond Wightman

Movement of cellulose synthase particles have so far been observed on the plant epidermis that are amenable to confocal imaging, yielding appreciable signal and resolution to observe small plasma membrane-localised particles. Presented here is a method, using airyscan confocal microscopy, that permits similar information to be obtained at depth within the developing protoxylem vessels of intact roots.

到目前为止,已经在植物表皮上观察到纤维素合酶颗粒的运动,这些颗粒适合共聚焦成像,产生可观的信号和分辨率来观察小的质膜局部颗粒。这里提出了一种方法,使用空气扫描共聚焦显微镜,允许在完整根的发育原木质部导管的深处获得类似的信息。
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引用次数: 0
Arabinogalactan proteins – Multifunctional glycoproteins of the plant cell wall 阿拉伯半乳聚糖蛋白——植物细胞壁的多功能糖蛋白
Q1 Immunology and Microbiology Pub Date : 2023-02-17 DOI: 10.1016/j.tcsw.2023.100102
Yingxuan Ma , Kim Johnson

Arabinogalactan-proteins (AGPs) are cell wall glycoproteins that make up a relatively small component of the extracellular matrix of plants yet have significant influence on wall mechanics and signalling. Present in walls of algae, bryophytes and angiosperms, AGPs have a wide range of functional roles, from signalling, cell expansion and division, embryogenesis, responses to abiotic and biotic stress, plant growth and development. AGPs interact with and influence wall matrix components and plasma membrane proteins to regulate developmental pathways and growth responses, yet the exact mechanisms remain elusive. Comprising a large gene family that is highly diverse, from minimally to highly glycosylated members, varying in their glycan heterogeneity, can be plasma membrane bound or secreted into the extracellular matrix, have members that are highly tissue specific to those with constitutive expression; all these factors have made it extremely challenging to categorise AGPs many qualities and roles. Here we attempt to define some key features of AGPs and their biological functions.

阿拉伯半乳聚糖蛋白(AGPs)是一种细胞壁糖蛋白,在植物细胞外基质中占相对较小的成分,但对细胞壁力学和信号传导有重大影响。AGPs存在于藻类、苔藓植物和被子植物的壁中,具有广泛的功能作用,从信号传导、细胞扩增和分裂、胚胎发生、对非生物和生物胁迫的反应、植物生长和发育。AGP与壁基质成分和质膜蛋白相互作用并影响其调节发育途径和生长反应,但其确切机制尚不清楚。包含一个高度多样化的大基因家族,从最低糖基化到高度糖基化的成员,其聚糖异质性各不相同,可以被质膜结合或分泌到细胞外基质中,其成员对具有组成型表达的成员具有高度组织特异性;所有这些因素使得对AGP的许多品质和角色进行分类变得极具挑战性。在这里,我们试图定义AGP的一些关键特征及其生物学功能。
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引用次数: 3
An update on xylan structure, biosynthesis, and potential commercial applications 木聚糖结构、生物合成和潜在商业应用的最新进展
Q1 Immunology and Microbiology Pub Date : 2023-01-28 DOI: 10.1016/j.tcsw.2023.100101
Thomas M. Curry , Maria J. Peña , Breeanna R. Urbanowicz
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引用次数: 11
A transferrin receptor’s guide to African trypanosomes 非洲锥虫的转铁蛋白受体指南
Q1 Immunology and Microbiology Pub Date : 2023-01-28 DOI: 10.1016/j.tcsw.2023.100100
Michael D. Urbaniak , Catarina Gadelha
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引用次数: 0
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