Revisiting the phosphotyrosine binding pocket of Fyn SH2 domain led to the identification of novel SH2 superbinders.

Protein engineering through directed evolution is an effective way to obtain proteins with novel functions with the potential applications as tools for diagnosis or therapeutics. Many natural proteins have undergone directed evolution in vitro in the test tubes in the laboratories worldwide, resulting in the numerous protein variants with novel or enhanced functions. we constructed here an SH2 variant library by randomizing 8 variable residues in its phosphotyrosine (pTyr) binding pocket. Selection of this library by a pTyr peptide led to the identification of SH2 variants with enhanced affinities measured by EC50. Fluorescent polarization was then applied to quantify the binding affinities of the newly identified SH2 variants. As a result, three SH2 variants, named V3, V13 and V24, have comparable binding affinities with the previously identified SH2 triple‐mutant superbinder. Biolayer Interferometry assay was employed to disclose the kinetics of the binding of these SH2 superbinders to the phosphotyrosine peptide. The results indicated that all the SH2 superbinders have two‐orders increase of the dissociation rate when binding the pTyr peptide while there was no significant change in their associate rates. Intriguingly, though binding the pTyr peptide with comparable affinity with other SH2 superbinders, the V3 does not bind to the sTyr peptide. However, variant V13 and V24 have cross‐reactivity with both pTyr and sTyr peptides. The newly identified superbinders could be utilized as tools for the identification of pTyr‐containing proteins from tissues under different physiological or pathophysiological conditions and may have the potential in the therapeutics.     

慢性乙型肝炎患者肠道菌群与肝脏生化指标的相关性

乙型肝炎病毒感染引起的慢性乙型肝炎 (Chronic hepatitis B,CHB) 是一种全球性流行疾病,严重时可引起肝功能衰竭,甚至发展成肝硬化和肝癌。也已发现CHB的发生和发展与肠道菌群的组成和结构的变化密切相关。为进一步探究肠道菌群结构与肝脏生化指标之间的联系,文中随机纳入14名CHB患者和11名健康对照者 (Control group,CN),分析其肝脏生化指标和肠道菌群的结构以及两者的相关性。结果发现CHB患者肝脏生化指标丙氨酸转氨酶 (Alanine transaminase,ALT)、总胆红素 (Total bilirubin,TBIL) 和γ-谷氨酰转移酶 (Gamma glutamyl transferase,GGT) 等的水平发生显著变化;普雷沃氏菌属Prevotella、劳特氏菌属Blautia、瘤胃球菌属Ruminococcus、Eubacterium eligens group等菌属失调;单形拟杆菌Bacteroides uniformis和Ruminococcus sp. 5_1_39BFAA可能与CHB患者的肝损伤相关,提示这些菌可能成为治疗CHB的新“切入点”。     

Crumbs proteins stabilize the cone mosaics of photoreceptors and improve vision in zebrafish

Although the unique organization of vertebrate cone mosaics was first described long ago,both their underlying molecular basis and physiological significance are largely unknown.Here,we demonstrate that Crumbs proteins,the key regulators of epithelial apical polarity,establish the planar cellular polarity of photoreceptors in zebrafish.Via heterophilic Crb2a-Crb2b interactions,the apicobasal polarity protein Crb2b restricts the asymmetric planar distribution of Crb2a in photoreceptors.The planar polarized Crumbs proteins thus balance intercellular adhesions and tension between photoreceptors,thereby stabilizing the geometric organization of cone mosaics.Notably,loss of Crb2b in zebrafish induces a nearsightedness-like phenotype in zebrafish accompanied by an elongated eye axis and impairs zebrafish visual perception for predation.These data reveal a detailed mechanism for cone mosaic homeostasis via previously undiscovered apical-planar polarity coordination and propose a pathogenic mechanism for nearsightedness.     

Dynamic Host Immune and Transcriptomic Responses to Respiratory Syncytial Virus Infection in a Vaccination-Challenge Mouse Model

Virologica Sinica - Respiratory syncytial virus (RSV) is the major cause of lower respiratory tract infections in children. Inactivated RSV vaccine was developed in the late 1960’s, but the...     

A Self-Biomineralized Novel Adenovirus Vectored COVID-19 Vaccine for Boosting Immunization of Mice

Virologica Sinica - SARS-CoV-2 has caused more than 3.8 million deaths worldwide, and several types of COVID-19 vaccines are urgently approved for use, including adenovirus vectored vaccines....     

聚多曲霉菌原生质体和液泡的制备及优化

原生质体的大量制备是研究原生质体转化、诱变、融合等技术的关键,液泡的制备对研究液泡中分解、转运有机物的特性具有重要意义,但目前分离技术还不够成熟.本研究从聚多曲霉菌菌丝中分离原生质体和液泡并对分离条件进行优化,以聚多曲霉菌DJ515-2菌丝为材料,探索不同因素对原生质体和液泡制备分离效果的影响.结果表明,聚多曲霉菌DJ515-2在菌龄42 h,以3％纤维素酶、1％蜗牛酶和3％的溶壁酶组成复合酶液,25℃下酶解4 h,原生质体达到最大产量,为5.167×105个/mL.同时,在此基础上裂解液泡的最优条件为pH7.5、1.0mol/L KCl、0.020％Triton X-10,其产量达2.63×105个/mL,产率为原生质体的50.8％,相比采用多元碱化合物诱导真菌原生质体裂解释放液泡的产率增加了 40％～45％.本研究为真菌和植物的各种原生质体技术及基于亚细胞层面的研究提供了材料基础.     

氧化甾醇结合蛋白相关蛋白家族的研究进展

膜脂是细胞膜的主要组分, 也是参与信号转导的重要信号分子。不同脂质分子在细胞膜上的不均等分布需要特殊类型的通道蛋白和运输蛋白来实现。氧化甾醇结合蛋白相关蛋白(ORPs)是一类非常保守的蛋白分子, 能够对磷脂酰肌醇和固醇等脂类分子进行识别并转运, 参与细胞中的许多生理过程, 包括信号转导、囊泡运输、脂类代谢和非囊泡运输等, 对于个体的生长发育具有重要作用。近几年, 关于ORPs在哺乳动物和酵母(Saccharomyces cerevisiae)中结构和功能的研究取得了一系列重要进展, 但在植物中相关研究尚少。该文综述了ORPs及其相关蛋白在哺乳动物、酵母和植物中的研究进展, 探讨了植物ORPs的结构及其与哺乳动物和酵母同源蛋白之间的进化关系, 并对植物ORPs未来的研究方向进行了展望。