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Keratinocyte and myeloid MCPIP1 have distinct roles in maintaining skin homeostasis 角质细胞和骨髓 MCPIP1 在维持皮肤稳态方面发挥着不同的作用。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-18 DOI: 10.1016/j.bbagen.2024.130671
Weronika Szukala , Agata Lichawska-Cieslar , Roza Zawada , Izabela Rumienczyk , Michal Mikula , Krzysztof Goryca , Jolanta Jura

The skin is a complex organ, and the intricate network between keratinocytes and immune cells is critical for ensuring skin function. Monocyte chemotactic protein-1-induced protein 1 (MCPIP1) is a ribonuclease that functions as a key negative modulator of inflammation. We previously reported that conditional deletion of MCPIP1 in keratinocytes (Mcpip1EKO) impairs skin integrity in adult mice. A similar phenotype was observed following the depletion of MCPIP1 in the myeloid compartment (Mcpip1MKO).

The aim of this study was to develop a keratinocyte and myeloid double-MCPIP1 knockout mouse model to clarify the specific roles of myeloid and epidermal MCPIP1 in skin biology.

Histological analyses indicated that the skin morphology changed after depletion of MCPIP1 in cells of myeloid origin as well as in keratinocytes. The thicknesses of the epidermal and subcutaneous fat layers increased in the mice with a loss of epidermal MCPIP1, whereas the loss of myeloid MCPIP1 had the opposite effect. In addition, both types of mice showed opposite responses to stimulation with 12-O-tetradecanoylphorbol-13-acetate. Transcriptomic profiling of whole-skin lysates revealed some common target transcripts in all the knockout mice. Further analyses revealed that distinct pathways are modulated following the loss of epidermal or myeloid MCPIP1. The skin morphology and inflammatory phenotype of keratinocyte and myeloid double-MCPIP1 knockout mice resembled those of mice with only keratinocyte-specific knockout of MCPIP1.

Overall, myeloid and epidermal MCPIP1 play important but distinct roles in the modulation of skin-related processes.

皮肤是一个复杂的器官,角质细胞和免疫细胞之间错综复杂的网络对确保皮肤功能至关重要。单核细胞趋化蛋白-1诱导蛋白1(MCPIP1)是一种核糖核酸酶,是炎症的关键负调控因子。我们以前曾报道过,有条件地缺失角质形成细胞中的 MCPIP1(Mcpip1EKO)会损害成年小鼠皮肤的完整性。在髓细胞中缺失 MCPIP1(Mcpip1MKO)后,也观察到了类似的表型。本研究的目的是建立一个角质细胞和髓细胞双 MCPIP1 基因敲除小鼠模型,以明确髓细胞和表皮 MCPIP1 在皮肤生物学中的特殊作用。组织学分析表明,髓源性细胞和角质形成细胞中的MCPIP1被耗尽后,皮肤形态发生了变化。表皮 MCPIP1 缺失的小鼠表皮层和皮下脂肪层厚度增加,而髓样 MCPIP1 缺失的小鼠则相反。此外,这两种类型的小鼠对 12-O-十四碳酰樟脑-13-乙酸酯的刺激表现出相反的反应。对全皮肤裂解液进行转录组分析发现,所有基因敲除小鼠都有一些共同的目标转录本。进一步的分析表明,表皮或骨髓 MCPIP1 缺失后,不同的通路会受到调节。角质细胞和髓质双 MCPIP1 基因敲除小鼠的皮肤形态和炎症表型与只敲除角质细胞特异性 MCPIP1 的小鼠相似。总之,髓质和表皮 MCPIP1 在调节皮肤相关过程中发挥着重要但不同的作用。
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引用次数: 0
Characterisation of biocondensate microfluidic flow using array-detector FCS 利用阵列检测器 FCS 对生物冷凝液微流体流动进行表征。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-17 DOI: 10.1016/j.bbagen.2024.130673
Stijn Dilissen , Pedro L. Silva , Anastasia Smolentseva , Tom Kache , Ronald Thoelen , Jelle Hendrix

Background

Biomolecular condensation via liquid-liquid phase separation (LLPS) is crucial for orchestrating cellular activities temporospatially. Although the rheological heterogeneity of biocondensates and the structural dynamics of their constituents carry critical functional information, methods to quantitatively study biocondensates are lacking. Single-molecule fluorescence research can offer insights into biocondensation mechanisms. Unfortunately, as dense condensates tend to sink inside their dilute aqueous surroundings, studying their properties via methods relying on Brownian diffusion may fail.

Methods

We take a first step towards single-molecule research on condensates of Tau protein under flow in a microfluidic channel of an in-house developed microfluidic chip. Fluorescence correlation spectroscopy (FCS), a well-known technique to collect molecular characteristics within a sample, was employed with a newly commercialised technology, where FCS is performed on an array detector (AD-FCS), providing detailed diffusion and flow information.

Results

The AD-FCS technology allowed characterising our microfluidic chip, revealing 3D flow profiles. Subsequently, AD-FCS allowed mapping the flow of Tau condensates while measuring their burst durations through the stationary laser. Lastly, AD-FCS allowed obtaining flow velocity and burst duration data, the latter of which was used to estimate the condensate size distribution within LLPS samples.

Conclusion

Studying biocondensates under flow through AD-FCS is promising for single-molecule experiments. In addition, AD-FCS shows its ability to estimate the size distribution in condensate samples in a convenient manner, prompting a new way of investigating biocondensate phase diagrams.

General significance

We show that AD-FCS is a valuable tool for advancing research on understanding and characterising LLPS properties of biocondensates.

背景:通过液-液相分离(LLPS)进行的生物分子凝聚对于协调细胞的时空活动至关重要。虽然生物凝结物的流变学异质性及其成分的结构动态具有重要的功能信息,但目前还缺乏定量研究生物凝结物的方法。单分子荧光研究可以深入了解生物缩合机制。遗憾的是,由于致密的凝聚物往往会在稀释的水环境中下沉,因此通过布朗扩散的方法来研究它们的性质可能会失败:方法:我们在自主研发的微流控芯片的微流控通道中对流动状态下的 Tau 蛋白凝聚物进行了单分子研究,迈出了第一步。荧光相关光谱(FCS)是一种收集样品内分子特征的著名技术,该技术采用了一种新的商业化技术,即在阵列探测器(AD-FCS)上进行荧光相关光谱分析,从而提供详细的扩散和流动信息:结果:AD-FCS 技术能够鉴定我们的微流控芯片,揭示三维流动曲线。随后,AD-FCS 可以绘制 Tau 凝聚物的流动图,同时通过固定激光测量其迸发持续时间。最后,AD-FCS 可以获得流速和迸发持续时间数据,后者用于估算 LLPS 样品中冷凝物的大小分布:结论:通过 AD-FCS 研究流动下的生物凝结物对于单分子实验很有前景。此外,AD-FCS 还能以一种便捷的方式估算冷凝物样品的尺寸分布,为研究生物冷凝物相图提供了一种新方法:我们的研究表明,AD-FCS 是推动了解和表征生物冷凝物 LLPS 特性研究的重要工具。
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引用次数: 0
Shotgun proteomics of thyroid carcinoma exosomes – Insight into the role of exosomal proteins in carcinogenesis and thyroid homeostasis 甲状腺癌外泌体的散射蛋白质组学研究--洞察外泌体蛋白质在癌变和甲状腺稳态中的作用。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-16 DOI: 10.1016/j.bbagen.2024.130672
Magdalena Surman , Magdalena Wilczak , Urszula Jankowska , Bożena Skupień-Rabian , Małgorzata Przybyło

Background

Transport of molecules via exosomes is one of the factors involved in thyroid cancer development, and transported molecules may serve as cancer biomarkers. The aim of the study was to characterize protein content of thyroid-derived exosomes and their functional effect exerted on recipient cells.

Methods

LC-MS/MS proteomics of exosomes released by FTC and 8305C thyroid carcinoma cell lines, and Nthy-ori 3–1 normal thyroid follicular cells was performed, followed by bioinformatic analysis and functional tests (wound healing and Alamar Blue assays).

Results

Exosomes from Nthy-ori 3–1 cells had the highest number of 1504 proteins, while in exosomes from thyroid carcinoma FTC and 8305C cells 730 and 1304 proteins were identified, respectively. For proteins uniquely found in FTC- and 8305C-derived exosomes, enriched cancer-related gene ontology categories included cell adhesion, positive regulation of cell migration, N-glycosylation, drug resistance, and response to NK/T cell cytotoxicity. Furthermore, through label-free quantification (that identified differentially expressed proteins) and comparison with The Human Protein Atlas database several potential diagnostic and/or prognostic biomarkers were indicated. Finally, exosomes from FTC and 8305C cells displayed ability to stimulate migratory properties of recipient Nthy-ori 3–1 cells. Additionally, 8305C-derived exosomes increased recipient cell viability.

Conclusions

Multiple proteins identified in thyroid cancer-derived exosomes have a direct link to thyroid cancer progression. Also, in functional tests exosomes enhanced growth and dissemination of non-transformed thyroid cells.

General significance

The obtained results expands the knowledge concerning the role of exosomal proteins in thyroid cancer and indicate potential biomarkers for further evaluation in clinical settings.

背景:通过外泌体转运分子是甲状腺癌发病的因素之一,转运的分子可作为癌症生物标志物。本研究旨在确定甲状腺外泌体的蛋白质含量及其对受体细胞的功能影响:方法:对FTC和8305C甲状腺癌细胞系以及Nthy-ori 3-1正常甲状腺滤泡细胞释放的外泌体进行LC-MS/MS蛋白质组学研究,然后进行生物信息分析和功能测试(伤口愈合和阿拉玛蓝试验):结果:Nthy-ori 3-1细胞的外泌体中有最多的1504个蛋白质,而甲状腺癌FTC细胞和8305C细胞的外泌体中分别发现了730个和1304个蛋白质。在FTC和8305C细胞外泌体中唯一发现的蛋白质中,与癌症相关的基因本体类别包括细胞粘附、细胞迁移的正向调节、N-糖基化、耐药性和对NK/T细胞细胞毒性的反应。此外,通过无标记定量(确定不同表达的蛋白质)以及与人类蛋白质图谱数据库的比较,还发现了一些潜在的诊断和/或预后生物标志物。最后,来自 FTC 和 8305C 细胞的外泌体显示出刺激受体 Nthy-ori 3-1 细胞迁移的能力。此外,8305C衍生的外泌体提高了受体细胞的活力:结论:在甲状腺癌外泌体中发现的多种蛋白质与甲状腺癌的进展有直接联系。结论:在甲状腺癌外泌体中发现的多种蛋白质与甲状腺癌的进展有直接联系。此外,在功能测试中,外泌体还能促进非转化甲状腺细胞的生长和扩散:研究结果拓展了有关外泌体蛋白在甲状腺癌中作用的知识,并为临床进一步评估提供了潜在的生物标记物。
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引用次数: 0
Mechanistic study of inhibitory peptides with SHP-1 in hypertonic environment for infection model 在高渗环境中用 SHP-1 对感染模型进行抑制肽的机制研究。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-10 DOI: 10.1016/j.bbagen.2024.130670
Shweta Khandibharad, Shailza Singh

Cutaneous Leishmaniasis, an infectious disease is globally the most prevalent form of leishmaniasis accounting for approximately 1 million cases every year as per world health organization. Infected individuals develop skin lesion which has been reported to be infiltrated by immune cells and parasite with high sodium accumulation creating hypertonic environment. In our work, we tried to mimic the hypertonic environment in virtual environment to study dynamicity of SHP-1 and NFAT5 along with their interactions through molecular dynamics simulation. We validated the SHP-1 and NFAT5 dynamics in infection and HSD conditions to study the impact of hypertonicity derived NFAT5 mediated response to L.major infection. We also evaluated our therapeutic peptides for their binding to SHP-1 and to form stable complex. Membrane stability with the peptides was analyzed to understand their ability to sustain mammalian membrane. We identified PepA to be a potential candidate to interact with SHP-1. Inhibition of SHP-1 through PepA to discern IL-10 and IL-12 reciprocity may be assessed in future and furnish us with a potential therapeutic molecule. HSD mice exhibited high pro-inflammatory response to L.major infection which resulted in reduced lesion size. Contrary to observations in HSD mice, infection model exhibited low pro-inflammatory response and increased lesion size with high parasite load. Thus, increase in NFAT5 expression and reduced SHP-1 expression may result in disease resolving effect which can be further studied through incorporation of synthetic circuit using PepA to modulate IL-10 and IL-12 reciprocity.

皮肤利什曼病是一种传染性疾病,是利什曼病在全球最常见的形式,据世界卫生组织统计,每年约有 100 万病例。据报道,受感染的个体会出现皮肤病变,皮肤被免疫细胞和寄生虫浸润,高钠积聚造成高渗环境。在我们的工作中,我们试图在虚拟环境中模拟高渗环境,通过分子动力学模拟研究 SHP-1 和 NFAT5 的动态及其相互作用。我们验证了 SHP-1 和 NFAT5 在感染和 HSD 条件下的动态,以研究高张力对 NFAT5 介导的大肠杆菌感染反应的影响。我们还评估了我们的治疗肽与 SHP-1 的结合并形成稳定的复合物。我们分析了多肽的膜稳定性,以了解它们维持哺乳动物膜的能力。我们发现 PepA 有可能与 SHP-1 相互作用。未来可能会评估通过 PepA 抑制 SHP-1,从而发现 IL-10 和 IL-12 之间的互作关系,并为我们提供一种潜在的治疗分子。HSD小鼠对L.major感染表现出较高的促炎反应,这导致了病变的缩小。与 HSD 小鼠的观察结果相反,感染模型表现出低促炎反应,寄生虫量高时病变面积增大。因此,NFAT5表达的增加和SHP-1表达的减少可能会导致疾病解决效应,这可以通过加入使用PepA调节IL-10和IL-12互作的合成回路来进一步研究。
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引用次数: 0
ROCK1 regulates glycolysis in pancreatic cancer via the c-MYC/PFKFB3 pathway ROCK1 通过 c-MYC/PFKFB3 通路调节胰腺癌中的糖酵解。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-10 DOI: 10.1016/j.bbagen.2024.130669

Background

Dysregulation of Rho-associated coiled coil-containing protein kinases (ROCKs) is involved in the metastasis and progression of various malignant tumors. However, how one of the isomers, ROCK1, regulates glycolysis in tumor cells is incompletely understood. Here, we attempted to elucidate how ROCK1 influences pancreatic cancer (PC) progression by regulating glycolytic activity.

Methods

The biological function of ROCK1 was analyzed in vitro by establishing a silenced cell model. Coimmunoprecipitation confirmed the direct binding between ROCK1 and c-MYC, and a luciferase reporter assay revealed the binding of c-MYC to the promoter of the PFKFB3 gene. These results were verified in animal experiments.

Results

ROCK1 was highly expressed in PC tissues and enriched in the cytoplasm, and its high expression was associated with a poor prognosis. Silencing ROCK1 inhibited the proliferation and migration of PC cells and promoted their apoptosis. Mechanistically, ROCK1 directly interacted with c-MYC, promoted its phosphorylation (Ser 62) and suppressed its degradation, thereby increasing the transcription of the key glycolysis regulatory factor PFKFB3, enhancing glycolytic activity and promoting PC growth. Silencing ROCK1 increased gemcitabine (GEM) sensitivity in vivo and in vitro.

Conclusions

ROCK1 promotes glycolytic activity in PC cells and promotes PC tumor growth through the c-MYC/PFKFB3 signaling pathway. ROCK1 knockdown can inhibit PC tumor growth in vivo and increase the GEM sensitivity of PC tumors, providing a crucial clinical therapeutic strategy for PC.

背景:Rho相关含线圈蛋白激酶(ROCKs)的失调与各种恶性肿瘤的转移和进展有关。然而,人们对异构体之一的 ROCK1 如何调节肿瘤细胞中的糖酵解尚不完全清楚。在此,我们试图阐明 ROCK1 如何通过调节糖酵解活性来影响胰腺癌(PC)的进展:方法:通过建立沉默细胞模型,在体外分析了 ROCK1 的生物学功能。免疫共沉淀证实了 ROCK1 与 c-MYC 之间的直接结合,荧光素酶报告实验显示了 c-MYC 与 PFKFB3 基因启动子的结合。这些结果在动物实验中得到了验证:结果:ROCK1在PC组织中高表达并富集于细胞质中,其高表达与预后不良有关。沉默 ROCK1 可抑制 PC 细胞的增殖和迁移,促进其凋亡。从机制上讲,ROCK1直接与c-MYC相互作用,促进其磷酸化(Ser 62)并抑制其降解,从而增加关键糖酵解调节因子PFKFB3的转录,增强糖酵解活性并促进PC生长。沉默ROCK1可提高体内和体外吉西他滨(GEM)的敏感性:结论:ROCK1能促进PC细胞的糖酵解活性,并通过c-MYC/PFKFB3信号通路促进PC肿瘤的生长。结论:ROCK1促进PC细胞的糖酵解活性,并通过c-MYC/PFKFB3信号通路促进PC肿瘤的生长。敲除ROCK1可抑制PC肿瘤在体内的生长,并提高PC肿瘤对吉西莫(GEM)的敏感性,从而为PC的临床治疗提供重要策略。
{"title":"ROCK1 regulates glycolysis in pancreatic cancer via the c-MYC/PFKFB3 pathway","authors":"","doi":"10.1016/j.bbagen.2024.130669","DOIUrl":"10.1016/j.bbagen.2024.130669","url":null,"abstract":"<div><h3>Background</h3><p>Dysregulation of Rho-associated coiled coil-containing protein kinases (ROCKs) is involved in the metastasis and progression of various malignant tumors. However, how one of the isomers, ROCK1, regulates glycolysis in tumor cells is incompletely understood. Here, we attempted to elucidate how ROCK1 influences pancreatic cancer (PC) progression by regulating glycolytic activity.</p></div><div><h3>Methods</h3><p>The biological function of ROCK1 was analyzed <em>in vitro</em> by establishing a silenced cell model. Coimmunoprecipitation confirmed the direct binding between ROCK1 and c-MYC, and a luciferase reporter assay revealed the binding of c-MYC to the promoter of the PFKFB3 gene. These results were verified in animal experiments.</p></div><div><h3>Results</h3><p>ROCK1 was highly expressed in PC tissues and enriched in the cytoplasm, and its high expression was associated with a poor prognosis. Silencing ROCK1 inhibited the proliferation and migration of PC cells and promoted their apoptosis. Mechanistically, ROCK1 directly interacted with c-MYC, promoted its phosphorylation (Ser 62) and suppressed its degradation, thereby increasing the transcription of the key glycolysis regulatory factor PFKFB3, enhancing glycolytic activity and promoting PC growth. Silencing ROCK1 increased gemcitabine (GEM) sensitivity <em>in vivo</em> and <em>in vitro</em>.</p></div><div><h3>Conclusions</h3><p>ROCK1 promotes glycolytic activity in PC cells and promotes PC tumor growth through the c-MYC/PFKFB3 signaling pathway. ROCK1 knockdown can inhibit PC tumor growth <em>in vivo</em> and increase the GEM sensitivity of PC tumors, providing a crucial clinical therapeutic strategy for PC.</p></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1868 10","pages":"Article 130669"},"PeriodicalIF":2.8,"publicationDate":"2024-07-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141598337","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Proteolytic cleavage of Golgi glycosyltransferases by SPPL3 and other proteases and its implications for cellular glycosylation SPPL3 和其他蛋白酶对高尔基糖基转移酶的蛋白水解作用及其对细胞糖基化的影响:(特刊标题:糖基转移酶的新方面)。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-09 DOI: 10.1016/j.bbagen.2024.130668

Glycosylation of proteins and lipids is of fundamental importance in multicellular eukaryotes. The vast diversity of glycan structures observed is generated in the Golgi apparatus by the concerted activity of >100 distinct enzymes, which include glycosyltransferases and other glycan-modifying enzymes. Well-known for decades, the majority of these enzymes is released from the Golgi apparatus and subsequently secreted into the extracellular space following endoproteolytic cleavage, but the underlying molecular mechanisms and the physiological implications have remained unexplored. This review will summarize our current knowledge of Golgi enzyme proteolysis and secretion and will discuss its conceptual implications for the regulation of cellular glycosylation and the organization of the Golgi apparatus. A particular focus will lie on the intramembrane protease SPPL3, which recently emerged as key protease facilitating Golgi enzyme release and has since been shown to affect a multitude of glycosylation-dependent physiological processes.

蛋白质和脂质的糖基化对多细胞真核生物至关重要。在高尔基体中,包括糖基转移酶和其他糖基修饰酶在内的 100 多种不同的酶协同作用,产生了多种多样的糖基结构。众所周知,这些酶的大部分都是从高尔基体释放出来的,随后经过内切蛋白裂解作用被分泌到细胞外空间,但其潜在的分子机制和生理影响仍有待探索。本综述将总结我们目前对高尔基体酶蛋白水解和分泌的认识,并讨论其对细胞糖基化调控和高尔基体组织的概念性影响。它是最近出现的促进高尔基体酶释放的关键蛋白酶,并被证明会影响多种依赖于糖基化的生理过程。
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引用次数: 0
Lectins of the Araceae family: Insights, distinctions, and future avenues—A three-decade investigation 天南星科植物的凝集素:洞察力、区别和未来途径--一项历时三十年的调查。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-05 DOI: 10.1016/j.bbagen.2024.130667
Emadeldin Hassan E. Konozy , Amina I. Dirar , Makarim Elfadil M. Osman

The Araceae family boasts >3000 species of flowering plants that thrive across the tropics. Among the focal points of study within this family are lectins, proteins with affinity for binding carbohydrates. This review endeavors to gather data gleaned from numerous studies conducted over the past three decades on lectins extracted from Araceae plants. Our examination spans their extraction and purification methods, their specific interactions with carbohydrates, their molecular structures, and various physicochemical characteristics. Furthermore, we investigated the biological activities of these lectins and investigated the outcomes of cloning their genes. Despite their apparent similarities, these lectins exhibit notable distinctions, particularly regarding their unique preferences in interacting with erythrocytes from animals and humans, their sugar affinities, the critical amino acids for their functionality, the molecular weights of their subunits and their respective topologies, and ultimately, their dimerization and 3D β-prism-II structure, which reportedly diverge from those observed in other GNA-related lectins. These discrepancies not only deepen our understanding of monocot lectins but also render these proteins inherently captivating. This review marks the inaugural attempt at consolidating almost all published reports on lectins from the Araceae family, with the aim of furnishing glycobiology scientists with essential insights into potential laboratory challenges, the characteristics of these lectins, and avenues for future research.

天南星科(Araceae)拥有超过 3000 种开花植物,它们在热带地区繁衍生息。凝集素是该科研究的重点之一,凝集素是一种具有结合碳水化合物亲和力的蛋白质。本综述试图收集过去三十年来从天南星科植物中提取的凝集素的大量研究数据。我们的研究涵盖了凝集素的提取和纯化方法、凝集素与碳水化合物的特殊相互作用、凝集素的分子结构以及各种理化特性。此外,我们还研究了这些凝集素的生物活性,并调查了克隆其基因的结果。尽管这些凝集素具有明显的相似性,但它们还是表现出了显著的差异,尤其是在与动物和人类红细胞相互作用的独特偏好、它们的糖亲和性、对它们的功能起关键作用的氨基酸、它们的亚基分子量和各自的拓扑结构,以及最终的二聚化和三维β-prism-II结构等方面,据报道,这些差异与在其他 GNA 相关凝集素中观察到的差异不同。这些差异不仅加深了我们对单子叶植物凝集素的了解,而且使这些蛋白质具有内在的吸引力。这篇综述首次尝试整合几乎所有已发表的有关芒柄蜡菊凝集素的报告,旨在为糖生物学科学家提供有关潜在的实验室挑战、这些凝集素的特征以及未来研究途径的重要见解。
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引用次数: 0
The malaria parasite egress protease SUB1 is activated through precise, plasmepsin X-mediated cleavage of the SUB1 prodomain 疟原虫出体蛋白酶 SUB1 是通过 plasmepsin X 介导的精确裂解 SUB1 原域激活的。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-07-03 DOI: 10.1016/j.bbagen.2024.130665
Chrislaine Withers-Martinez , Roger George , Sarah Maslen , Létitia Jean , Fiona Hackett , Mark Skehel , Michael J. Blackman

Background

The malaria parasite Plasmodium falciparum replicates within red blood cells, then ruptures the cell in a process called egress in order to continue its life cycle. Egress is regulated by a proteolytic cascade involving an essential parasite subtilisin-like serine protease called SUB1. Maturation of SUB1 initiates in the parasite endoplasmic reticulum with autocatalytic cleavage of an N-terminal prodomain (p31), which initially remains non-covalently bound to the catalytic domain, p54. Further trafficking of the p31-p54 complex results in formation of a terminal p47 form of the SUB1 catalytic domain. Recent work has implicated a parasite aspartic protease, plasmepsin X (PMX), in maturation of the SUB1 p31-p54 complex through controlled cleavage of the prodomain p31.

Methods

Here we use biochemical and enzymatic analysis to examine the activation of SUB1 by PMX.

Results

We show that both p31 and p31-p54 are largely dimeric under the relatively acidic conditions to which they are likely exposed to PMX in the parasite. We confirm the sites within p31 that are cleaved by PMX and determine the order of cleavage. We find that cleavage by PMX results in rapid loss of the capacity of p31 to act as an inhibitor of SUB1 catalytic activity and we directly demonstrate that exposure to PMX of recombinant p31-p54 complex activates SUB1 activity.

Conclusions

Our results confirm that precise, PMX-mediated cleavage of the SUB1 prodomain activates SUB1 enzyme activity.

General significance

Our findings elucidate the role of PMX in activation of SUB1, a key effector of malaria parasite egress.

背景:恶性疟原虫在红细胞内复制,然后在一个称为 "出路 "的过程中破裂细胞,以继续其生命周期。细胞外排受蛋白水解级联的调控,其中涉及一种名为 SUB1 的重要寄生虫类丝氨酸蛋白酶。SUB1 在寄生虫内质网中的成熟始于 N 端原域(p31)的自催化裂解,该原域最初与催化域 p54 保持非共价结合。p31-p54 复合物的进一步转移导致形成 SUB1 催化域的末端 p47 形式。最近的研究表明,寄生虫天冬氨酸蛋白酶 plasmepsin X(PMX)通过控制原域 p31 的裂解,参与了 SUB1 p31-p54 复合物的成熟:结果:我们发现,在寄生虫体内,p31 和 p31-p54 很可能暴露于 PMX 的相对酸性条件下,它们在很大程度上都是二聚体。我们确认了 p31 中被 PMX 裂解的位点,并确定了裂解的顺序。我们发现,PMX 的裂解会导致 p31 迅速丧失作为 SUB1 催化活性抑制剂的能力,而且我们直接证明,重组 p31-p54 复合物暴露于 PMX 会激活 SUB1 的活性:我们的研究结果证实,PMX 介导的精确裂解 SUB1 原域可激活 SUB1 酶的活性:我们的研究结果阐明了 PMX 在激活 SUB1 过程中的作用,SUB1 是疟原虫出体的关键效应因子。
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引用次数: 0
YAP1-mediated dysregulation of ACE-ACE2 activity augments cardiac fibrosis upon induction of hyperglycemic stress YAP1 介导的 ACE-ACE2 活性失调会在诱导高血糖应激时加重心脏纤维化。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-30 DOI: 10.1016/j.bbagen.2024.130666
Arunima Mondal , Shreya Das , Madhuchhanda Das , Santanu Chakraborty , Arunima Sengupta

Background

Diabetic stress acts on the cardiac tissue to induce cardiac hypertrophy and fibrosis. Diabetes induced activated renin angiotensin system (RAS) has been reported to play a critical role in mediating cardiac hypertrophy and fibrosis. Angiotensin converting enzyme (ACE) in producing Angiotensin-II, promotes cardiomyocyte hypertrophy and fibrotic damage. ACE2, a recently discovered molecule structurally homologous to ACE, has been reported to be beneficial in reducing the effect of RAS driven pathologies.

Methods

In vivo diabetic mouse model was used and co-labelling immunostaining assay have been performed to analyse the fibrotic remodeling and involvement of associated target signaling molecules in mouse heart tissue. For in vitro analyses, qPCR and western blot experiments were performed in different groups for RNA and protein expression analyses.

Results

Fibrosis markers were observed to be upregulated in the diabetic mouse heart tissue as well as in high glucose treated fibroblast and cardiomyocyte cells. Hyperglycemia induced overexpression of YAP1 leads to increased expression of β-catenin (CTNNB1) and ACE with downregulated ACE2 expression. The differential expression of ACE/ACE2 promotes TGFB1-SMAD2/3 pathway in the hyperglycemic cardiomyocyte and fibroblast resulting in increased cardiac fibrotic remodeling.

Conclusion

In the following study, we have reported YAP1 modulates the RAS signaling pathway by inducing ACE and inhibiting ACE2 activity to augment cardiomyocyte hypertrophy and fibrosis in hyperglycemic condition. Furthermore, we have shown that hyperglycemia induced dysregulation of ACE-ACE2 activity by YAP1 promotes cardiac fibrosis through β-catenin/TGFB1 dependent pathway.

背景:糖尿病压力作用于心脏组织,诱发心脏肥大和纤维化。据报道,糖尿病诱导的活化肾素血管紧张素系统(RAS)在介导心脏肥大和纤维化方面起着关键作用。血管紧张素转换酶(ACE)产生血管紧张素-II,促进心肌细胞肥大和纤维化损伤。据报道,最近发现的与血管紧张素转换酶结构同源的分子 ACE2 有助于减轻 RAS 驱动的病变的影响:方法:使用体内糖尿病小鼠模型和共标记免疫染色法分析小鼠心脏组织的纤维化重塑和相关靶信号分子的参与。在体外分析中,对不同组的 RNA 和蛋白质表达进行了 qPCR 和 Western 印迹实验:结果:在糖尿病小鼠心脏组织以及经高糖处理的成纤维细胞和心肌细胞中观察到纤维化标志物上调。高血糖诱导的 YAP1 过表达导致β-catenin(CTNNB1)和 ACE 的表达增加,而 ACE2 的表达下调。ACE/ACE2的不同表达促进了高血糖心肌细胞和成纤维细胞中的TGFB1-SMAD2/3通路,导致心脏纤维化重塑增加:在接下来的研究中,我们报道了 YAP1 通过诱导 ACE 和抑制 ACE2 的活性来调节 RAS 信号通路,从而在高血糖条件下促进心肌细胞肥大和纤维化。此外,我们还发现,高血糖诱导的 YAP1 对 ACE-ACE2 活性的失调会通过 β-catenin/TGFB1 依赖性途径促进心脏纤维化。
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引用次数: 0
The Chinese medaka (Oryzias sinensis) dmrt1 gene converts females to males in medaka (Oryzias latipes) 中国青鳉(Oryzias sinensis)的 dmrt1 基因可将雌性转化为雄性。
IF 2.8 3区 生物学 Q3 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-06-26 DOI: 10.1016/j.bbagen.2024.130664
Lei Chen , Yan Huang , Qi-Hua Pan , Meng-Yang Wang , Jing-Jie Liang , Tian-Sheng Chen

Background

Chinese medaka (Oryzias sinensis) is widely distributed in freshwater rivers in China. Similar to the medaka (Oryzias latipes), Chinese medaka has the characteristics of small size, rapid reproductive cycle, and strong adaptability, which makes it suitable as a model organism for studies in basic biology and environmental toxicology. Chinese medaka exhibits distinct sexual dimorphism. However, due to the lack of complete genomic information, the regulation of sex determination and differentiation-related genes in Chinese medaka remains unclear.

Methods

Chinese medaka dmrt1 (Osdmrt1) was cloned by PCR, and transgenic individuals of medaka [Tg(CMV:Osdmrt1)] overexpressing Osdmrt1 were generated to investigate the role of Osdmrt1 in sex determination. Western blot was used to validate the integration of the Osdmrt1 into the medaka genome. Tissue sectioning and HE staining were used to identify Tg(CMV:Osdmrt1) physiological gender and phenotype. qRT-PCR was used to analyze the expression of gonad-specific genes.

Results

Osdmrt1 was cloned and identified, and it shared similar evolutionary relationships with medaka dmrt1. Tg(CMV:Osdmrt1) exhibited partial sex reversal from female to male in the F2 generation, with genetically female individuals developing testes and producing functional sperm. Additionally, the secondary sexual characteristics of the transgenic females also changed to males.

Conclusion

The Chinese medaka dmrt1 gene could convert females to males in medaka.

General significance: These results not only elucidate the function of Chinese medaka dmrt1, but also accumulate knowledge for studying the function of economically important fish genes in model fish by transgenic technology.

背景中国青鳉广泛分布于中国的淡水河流中。与青鳉相似,中国青鳉具有体型小、繁殖周期快、适应性强等特点,适合作为基础生物学和环境毒理学研究的模式生物。中国青鳉具有明显的性二型。然而,由于缺乏完整的基因组信息,中国青鳉性别决定和分化相关基因的调控仍不清楚:方法:利用PCR技术克隆了中国青鳉的dmrt1(Osdmrt1),并产生了过表达Osdmrt1的转基因青鳉个体[Tg(CMV:Osdmrt1)],以研究Osdmrt1在性别决定中的作用。用 Western blot 验证了 Osdmrt1 与青鳉基因组的整合。用组织切片和HE染色鉴定Tg(CMV:Osdmrt1)的生理性别和表型:结果:克隆并鉴定了Osdmrt1,它与青鳉Dmrt1具有相似的进化关系。Tg(CMV:Osdmrt1)在F2代表现出从雌性到雄性的部分性别逆转,基因上为雌性的个体发育出睾丸并产生功能性精子。此外,转基因雌性个体的第二性征也变为雄性:结论:中国青鳉 dmrt1 基因可使雌性青鳉转为雄性:这些结果不仅阐明了中国青鳉 dmrt1 的功能,而且为利用转基因技术研究经济鱼类功能基因在模式鱼类中的功能积累了知识。
{"title":"The Chinese medaka (Oryzias sinensis) dmrt1 gene converts females to males in medaka (Oryzias latipes)","authors":"Lei Chen ,&nbsp;Yan Huang ,&nbsp;Qi-Hua Pan ,&nbsp;Meng-Yang Wang ,&nbsp;Jing-Jie Liang ,&nbsp;Tian-Sheng Chen","doi":"10.1016/j.bbagen.2024.130664","DOIUrl":"10.1016/j.bbagen.2024.130664","url":null,"abstract":"<div><h3>Background</h3><p>Chinese medaka (<em>Oryzias sinensis</em>) is widely distributed in freshwater rivers in China. Similar to the medaka (<em>Oryzias latipes</em>), Chinese medaka has the characteristics of small size, rapid reproductive cycle, and strong adaptability, which makes it suitable as a model organism for studies in basic biology and environmental toxicology. Chinese medaka exhibits distinct sexual dimorphism. However, due to the lack of complete genomic information, the regulation of sex determination and differentiation-related genes in Chinese medaka remains unclear.</p></div><div><h3>Methods</h3><p>Chinese medaka <em>dmrt1</em> (Os<em>dmrt1</em>) was cloned by PCR, and transgenic individuals of medaka [Tg(CMV:Os<em>dmrt1</em>)] overexpressing Os<em>dmrt1</em> were generated to investigate the role of Os<em>dmrt1</em> in sex determination. Western blot was used to validate the integration of the Os<em>dmrt1</em> into the medaka genome. Tissue sectioning and HE staining were used to identify Tg(CMV:Os<em>dmrt1</em>) physiological gender and phenotype. qRT-PCR was used to analyze the expression of gonad-specific genes.</p></div><div><h3>Results</h3><p>Os<em>dmrt1</em> was cloned and identified, and it shared similar evolutionary relationships with medaka <em>dmrt1</em>. Tg(CMV:Os<em>dmrt1</em>) exhibited partial sex reversal from female to male in the F2 generation, with genetically female individuals developing testes and producing functional sperm. Additionally, the secondary sexual characteristics of the transgenic females also changed to males.</p></div><div><h3>Conclusion</h3><p>The Chinese medaka <em>dmrt1</em> gene could convert females to males in medaka.</p><p><em><strong>General significance:</strong></em> These results not only elucidate the function of Chinese medaka <em>dmrt1</em>, but also accumulate knowledge for studying the function of economically important fish genes in model fish by transgenic technology.</p></div>","PeriodicalId":8800,"journal":{"name":"Biochimica et biophysica acta. General subjects","volume":"1868 9","pages":"Article 130664"},"PeriodicalIF":2.8,"publicationDate":"2024-06-26","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141465913","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
期刊
Biochimica et biophysica acta. General subjects
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