鳙鳃凝集素的分离纯化及理化性质的研究     

盘赛昆  汤坚  顾小红  杨超 《水产科学》2009,28(12)

利用DEAE-Sepharose FF离子交换和Sephacryl S-200 HR及Superdex 200 10/300 GL分子筛层析技术,从鳙鳃组织中分离纯化到一种岩藻糖专一的凝集素,命名为GANL.在还原SDS-PAGE电泳上显示单一蛋白染色带,其亚基相对分子质量为37 kD.经Superdex 200凝胶过滤层析测得其天然相对分子质量为220 kD.GANL的中性糖含量为13.4%.因此,GANL作为一种由相同亚基组成多亚基糖蛋白.GANL对兔红细胞有专一的凝集活性,其凝集活性不依赖Ca~(2+).在被测的单糖、双糖及糖蛋白中,仅岩藻糖能抑制其凝集活性.氨基酸分析表明GANL的Asp、Glu、Leu、Val、Lys的含量较高,Cys-S的含量为0.81%.GANL的最适pH为8～9;具有很高的热稳定性,最适温度为50 ℃.  相似文献   

日本沼虾C型凝集素结构域家族3的cDNA克隆、原核表达和定位分析     

孙盛明  傅洪拓  宣富君  戈贤平  朱健  吴旭干 《水产学报》2019,43(11):2317-2326

C型凝集素(C-type lectin)是一类能与糖类结合的非抗体的蛋白质或糖蛋白家族,为了研究C型凝集素基因在日本沼虾组织分布、细胞定位和细菌感染过程中的表达情况,本研究应用cDNA末端快速克隆(rapid-amplification of cDNA ends,RACE)技术首次克隆了日本沼虾C型凝集素结构域家族3基因(MnLec3)的全长序列,通过实时荧光定量PCR(qRT-PCR)分析MnLec3基因在不同组织、细菌感染后不同时间的表达水平,Western blot和免疫荧光分别分析蛋白的表达水平和细胞定位。结果显示,MnLec3基因cDNA全长1 357 bp,包括125 bp的5′末端非翻译区(UTR)、1 026 bp的开放阅读框(ORF)和206 bp的3′UTR,其中开放阅读框编码341个氨基酸。氨基酸序列比对显示,日本沼虾MnLec3基因含有保守钙结合点(Met 1-Glu17)和糖识别结构域(CRD)。同源性分析结果显示,MnLec3与罗氏沼虾C型凝集素3相似度较高;邻接法(Neighbor-Joining,NJ)进化树分析结果显示,MnLec3与其他甲壳动物C型凝集素聚为一支。通过构建原核表达载体获得体外重组蛋白rMnLec3,并将纯化重组蛋白免疫大鼠获得抗血清,免疫荧光结果显示,绿色荧光信号主要在肝胰腺细胞核中表达。qRT-PCR结果显示,MnLec3在日本沼虾所检测组织中均表达,其中肝胰腺中表达量最高,血细胞次之;与对照组相比,在嗜水气单胞菌刺激12～48 h时MnLec3表达量显著升高,48 h表达量最高,Western blot分析结果显示,MnLec3蛋白表达丰度与基因表达模式基本相似,提示克隆得到的MnLec3参与日本沼虾抵御细菌入侵的免疫过程。  相似文献   

Algal and Cyanobacterial Lectins and Their Antimicrobial Properties     

Jos Abel Fernndez Romero  María Gabriela Paglini  Christine Priano  Adolfina Koroch  Yoel Rodríguez  James Sailer  Natalia Teleshova 《Marine drugs》2021,19(12)

Lectins are proteins with a remarkably high affinity and specificity for carbohydrates. Many organisms naturally produce them, including animals, plants, fungi, protists, bacteria, archaea, and viruses. The present report focuses on lectins produced by marine or freshwater organisms, in particular algae and cyanobacteria. We explore their structure, function, classification, and antimicrobial properties. Furthermore, we look at the expression of lectins in heterologous systems and the current research on the preclinical and clinical evaluation of these fascinating molecules. The further development of these molecules might positively impact human health, particularly the prevention or treatment of diseases caused by pathogens such as human immunodeficiency virus, influenza, and severe acute respiratory coronaviruses, among others.  相似文献   

多子小瓜虫滋养体糖蛋白残基的种类和分布研究     

赖晓健  熊静  张哲  李忠琴 《水产科学》2016,(6):713-717

应用伴刀豆凝集素A、麦胚凝集素、大豆凝集素和荆豆凝集素Ⅰ,检测寄生于金鱼皮肤、鳃和鳍上的多子小瓜虫滋养体糖蛋白残基的种类和分布。研究结果发现,伴刀豆凝集素A和麦胚凝集素免疫阳性染色在金鱼皮肤、鳃和鳍寄生的滋养体上均有分布,麦胚凝集素免疫阳性染色强于伴刀豆凝集素A,未见有大豆凝集素和荆豆凝集素Ⅰ免疫阳性染色。鳃上寄生的滋养体伴刀豆凝集素A免疫阳性染色最强,皮肤次之,鳍最弱。鳍上寄生的滋养体麦胚凝集素免疫阳性染色最强,皮肤次之,鳃最弱。研究结果表明,滋养体有单糖D-甘露糖和D-葡萄糖,以及氨基衍生物乙酰氨基葡萄糖。寄生于鳃上的滋养体D-甘露糖和D-葡萄糖可能较多,寄生在鳍上的滋养体乙酰氨基葡萄糖可能较多。  相似文献   

Entry Inhibition of Influenza Viruses with High Mannose Binding Lectin ESA-2 from the Red Alga Eucheuma serra through the Recognition of Viral Hemagglutinin     

Yuichiro Sato  Kinjiro Morimoto  Takanori Kubo  Takemasa Sakaguchi  Akira Nishizono  Makoto Hirayama  Kanji Hori 《Marine drugs》2015,13(6):3454-3465

Lectin sensitivity of the recent pandemic influenza A virus (H1N1-2009) was screened for 12 lectins with various carbohydrate specificity by a neutral red dye uptake assay with MDCK cells. Among them, a high mannose (HM)-binding anti-HIV lectin, ESA-2 from the red alga Eucheuma serra, showed the highest inhibition against infection with an EC₅₀ of 12.4 nM. Moreover, ESA-2 exhibited a wide range of antiviral spectrum against various influenza strains with EC₅₀s of pico molar to low nanomolar levels. Besides ESA-2, HM-binding plant lectin ConA, fucose-binding lectins such as fungal AOL from Aspergillus oryzae and AAL from Aleuria aurantia were active against H1N1-2009, but the potency of inhibition was of less magnitude compared with ESA-2. Direct interaction between ESA-2 and a viral envelope glycoprotein, hemagglutinin (HA), was demonstrated by ELISA assay. This interaction was effectively suppressed by glycoproteins bearing HM-glycans, indicating that ESA-2 binds to the HA of influenza virus through HM-glycans. Upon treatment with ESA-2, no viral antigens were detected in the host cells, indicating that ESA-2 inhibited the initial steps of virus entry into the cells. ESA-2 would thus be useful as a novel microbicide to prevent penetration of viruses such as HIV and influenza viruses to the host cells.  相似文献   

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways     

Imtiaj Hasan  Shigeki Sugawara  Yuki Fujii  Yasuhiro Koide  Daiki Terada  Naoya Iimura  Toshiyuki Fujiwara  Keisuke G. Takahashi  Nobuhiko Kojima  Sultana Rajia  Sarkar M. A. Kawsar  Robert A. Kanaly  Hideho Uchiyama  Masahiro Hosono  Yukiko Ogawa  Hideaki Fujita  Jiharu Hamako  Taei Matsui  Yasuhiro Ozeki 《Marine drugs》2015,13(12):7377-7389

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.  相似文献   

Sea bass Dicentrarchus labrax (L.) bacterial infection and confinement stress acts on F‐type lectin (DlFBL) serum modulation          下载免费PDF全文

M G Parisi  G Benenati  M Cammarata 《Journal of fish diseases》2015,38(11):967-976

The F‐lectin, a fucose‐binding protein found from invertebrates to ectothermic vertebrates, is the last lectin family to be discovered. Here, we describe effects of two different types of stressors, bacterial infection and confinement stress, on the modulation of European sea bass Dicentrarchus labrax (L.) F‐lectin (DlFBL), a well‐characterized serum opsonin, using a specific antibody. The infection of the Vibrio alginolyticus bacterial strain increased the total haemagglutinating activity during the 16‐day testing period. The DlFBL value showed an upward regulation on the first, second and last days and underwent a slight downward regulation 4 days post‐challenge. In contrast, the effect of confinement and density stress showed a decrease in the plasma concentration of lectin, ranging from 50% to 60% compared with the control. The modulation of DlFBL is in line with the hypothesis that humoral lectins could be involved and recruited in the initial recognition step of the inflammation, which leads to agglutination, and the activation of mechanisms responsible for killing of the pathogens.  相似文献   

蒜头果凝集素的提取及血凝效果研究   总被引：1，自引：1，他引：0  

袁燕袁再美  岑晓江程霞 《中国农学通报》2011,27(21):144-146

从蒜头果种仁中提取凝集素,用20%、40%、60%、80%、100%饱和度硫酸铵分级沉淀,分别用鸡、兔的红细胞悬液检测其凝血活性。结果表明,80%以下硫酸铵沉淀物均可与鸡、兔的红细胞悬液发生凝集反应,100%的硫酸铵沉淀物则没有凝集反应,说明用硫酸铵沉淀到80%饱和度以后,蒜头果凝集素即可完全沉淀出来。此结果证明蒜头果中含有凝集素,为蒜头果凝集素的开发利用奠定了基础。  相似文献   

不同地区马铃薯凝集素含量与活性的测定分析   总被引：1，自引：0，他引：1  

王宝枝 《湖北农业科学》2009,48(4)

植物凝集素是一个超级蛋白质家族,能凝集细胞.不同植物凝集素含量和活性均不相同,即使同一品种植物由于所生长的地区环境不同,其含量和活性也各不相同.本试验对两个不同地区的白瓤马铃薯就此两方面进行了初步研究.结果发现.东北马铃薯的凝集素含量和活性远大于山东马铃薯.  相似文献   

不同马铃薯品种凝集素含量和活性比较     

王宝枝 《贵州农业科学》2009,37(7)

植物凝集素是一类具有高度特异性的糖结合活性的蛋白,能凝集细胞.不同植物凝集素含量和活性均不相同,即使同一种植物由于品种不同,其含量和活性也各不相同,就此两方面,试验首先通过葡聚糖凝胶层析法和分光光度法得出黄瓤马铃薯凝集素含量多于白瓤马铃薯,继而通过血凝平板试验得出同等条件下黄瓤马铃薯凝集素活性也大于白瓤的.表明,黄瓤马铃薯凝集素含量和活性均大于白瓤马铃薯.  相似文献