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Characterization and Phylogenetic Analysis of lectin Gene cDNA Isolated from Sea Cucumber (Apostichopus japonicus) Body Wall

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Journal of Ocean University of China 2017年第6期16卷 1148-1156页

作者： XUE Zhuang LI Hui LIU Yang ZHOU Wei SUN Jing WANG Xiuli College of Fisheries and Life Science Dalian Ocean University Dalian 116023 P. R. China Liaoning Province Academy of Analytic Science Shenyang 110015 P. R. China

As a ‘living fossil' of species origin and ‘rich treasure' of food and nutrition development, sea cucumber has received a lot of attentions from researchers. The cDNA library construction and EST sequencing of blood had been conducted previously in our lab. The bioinformatic analysis provided a gene fragment which is highly homologous with the genes of lectin family, named AjL(Apostichopus japonicus lectin). To characterize and determine the phylogeny of AjL genes in early evolution, we isolated a fulllength cDNA of lectin gene from the body wall of A. japonicus. The open reading frame of this gene contained 489 bp and encoded a 163 amino acids secretory protein being homologous to lectins of mammals and aquatic organisms. The deduced protein included a lectin-like domain. SDS-PAGE analysis showed that AjL migrated as a specific band(about 36.09 kDa under reducing), and agglutinated against rabbit red blood cells. AjL was similar to chain A of CEL-IV in space structure. We predicted that AjL may play the same role of CEL-IV. Our results suggested that more than one lectin gene functioned in sea cucumber and most of other species, which was fused by uncertain sequences during the evolution and encoded different proteins with diverse functions. Our findings provided the insights into the function and characteristics of lectin genes invertebrates. The results will also be helpful for the identification and structural, functional, and evolutionary analyses of lectin genes.

关键词： sea cucumber agglutination activity lectin space structure

Effect of Pinellia ternata lectin on Membrane Currents of Mouse Motor Nerve Terminals

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Science China Chemistry 1994年第4期37卷 448-453页

作者：施玉樑徐幼芬张辉 Shanghai Institute of Physiology Academia Sinica Shanghai 200031 PRC

Pinellia ternata lectin (PTL) extracted from the fresh juice of rhizome of pinellia ternata used as a traditional Chinese medicine facilitated the quantal release of acetylcholine (ACh) in the mouse motor nerve terminals and formed cation channels in artificial lipid bilayer. Here we report the action of PTL on presynaptic membrane currents of motor nerve *** experiments were performed on the intercostal nerve triangularis sterni muscle preparations. By means of the perineurial recording, the effects of PTL on the sodium current in the preterminal part , three potassium currents and two calcium currents generated from the nerve terminals were investigated. The results show that PTL increases voltage-dependent fast Ca2+ current (ICa,f), Na+ current (INa) and Ca2+-acti-vated K+ current (IK,Ca) without action on either the voltage-dependent fast K+ current (IK,f) or the slow K+ current (IK,S). These effects are irreversible, but can be reversed by mannan, the specific binding sugar for *** total action of PTL can be considered to raise [Ca2+], by promoting Ca2+ influx, and then facilitate the neurotransmitter release. To our knowledge, this is the first report about the influence of lectin on presynaptic membrane currents at mammalian motor nerve terminals.

关键词： Pinellia ternata lectin motor nerve terminal peYineurial recording membrane currents.

cDNA cloning of a novel lectin that induce cell apoptosis from Artocarpus hypargyreus

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Chinese Journal of Natural Medicines 2021年第2期19卷 81-89页

作者： LUO Yu ZENG Lin-Jie LIU Xiao-Qin LI Lu ZENG Qi-Yan Department of Biochemistry and Molecular Biology Guangxi Medical UniversityNanning 530021China Department of Orthopaedics Chinese and Western Orthopaedics Hospital of GuigangGuigang 537100China

We isolated a novel lectin(AHL)from Artocarpus hypargyreus Hance and showed its immunomodulatory *** this study,the amino acid sequence of AHL was determined by cDNA *** cDNA(875bp)contains a 456-bp open reading frame(ORF),which encodes a protein with 151 amino *** is a new member of jacalin-related lectin family(JRLs),which share high sequence similarities to KM+and Morniga M,and contain the conserved carbohydrate binding *** antitumor activity of AHL was also explored using Jurkat T cell *** exhibits a strong binding affinity to cell membrane,which can be effectively inhibited by methyl-α-*** inhibits cell proliferation in a time-and dose-dependent manner through apoptosis,evidenced by morphological changes,phosphatidylserine externalization,poly ADP-ribose polymerase(PARP)cleavage,Bad and Bax up-regulation,and caspase-3 *** further showed that the activation of ERK and p38 signaling pathways is involved for the pro-apoptotic effect of AHL.

关键词： lectin cDNA cloning Apoptosis Antitumor drug Artocarpus hypargyreus

Cloning and molecular characterization of a novel lectin gene from Pinellia ternata

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Cell Research 2003年第4期13卷 301-308页

作者： JIAN HONG YAO, XIU YUN ZHAO, ZHI HUA LIAO, JUAN LIN, ZHONG HAI CHEN, FEI CHEN, JUN SONG, XIAO FEN SUN, KE XUAN TANG State Key Laboratory of Genetic Engineering, School of Life Sciences, Morgan- Tan International Center for Life Sciences, Fudan-SJTU-Nottingham Plant Biotechnology R and D Center, Fudan University, Shanghai 200433, ChinaFudan-SJTU-Nottingham Plant Biotechnology R and D Center, Plant Biotechnology Research Center, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200030, China State Key Laboratory of Genetic Engineering School of Life Sciences Morgan-Tan International Center for Life Sciences Fudan-SJTU-Nottingham Plant Biotechnology R and D Center Fudan University Shanghai 200433 China

The full-length cDNA of Pinellia ternata agglutinin (PTA) was cloned from inflorescences using RACE PCR. Through comparative analysis of PTA gene (pta) and its deduced amino acid sequence with those of other Araceae species, pta was found to encode a precursor lectin with signal peptide and to have extensive homology with those of other Araceae species. PTA was a heterotetrameric mannose-binding lectin with three mannose-binding boxes like lectins from other Araceae and Amaryllidaceae species. Southern blot analysis of the genomic DNA revealed that pta belonged to a low-copy gene family. Northern blot analysis demonstrated that pta constitutively expressed in various plant tissues including root, leaf, stem and inflorescence. The pta cDNA sequence encoding for mature PTA protein was cloned into pET-32a plasmid and the resulting plasmid, pET-32a-PTA containing Trx-PTA fusion protein, was investigated for the expression in E. coli BL21. SDS-PAGE gel analysis showed that the Trx-PTA fusion protein was succ

关键词： araceae cDNA cloning lectin Pinellia ternata RACE-PCR.

Cation Selectivity of Channels Formed at Planar Lipid Bilayer by Pinellia ternata lectin

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Science China Chemistry 1994年第5期37卷 547-556页

作者：张辉施玉醻 Shanghai Institute of Physiology Academia Sinica Shanghai 200031 PRC

By means of the two-compartment system. PTL-channels, the first lectin channels formed on planar lipid bilayers by Pinellia ternata lectin (PTL) have been studied. The results show (i) PTL-channels are voltage-independent and have apparent subunits; (ii) in 50 mmol/L KC1 and 25 mmol/L BaCl2 solutions, a single channel has unit conductance of 21 pS and 42 pS, respectively; (iii) the channel exhibits a slightly higher permeability to divalent than monovalent cation (PBa/PK=4.1), and (iv) the selectivity among divalent or monovalent cations is poor. The cation selectivity sequence for the channel will follow PBa(7. 0)-Psr (6. 4)~PMg(6. 4)>PK(1. 7)>PN.(1. 0)~PLi(l. 0).Moreover, these data also give explanation to the facilitatory action of PTL on the release of acetyl-choline from motor nerve terminals.

关键词： Pinellia ternata Briet lectin lipid bilayer single-channel recording cation selectivity.

Comparative analysis of C-type lectin domain proteins in the ghost moth, Thitarodes xiaojinensis (Lepidoptera: Hepialidae)

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Insect Science 2019年第3期26卷 453-465页

作者： Qian Meng Ji-Hong Zhang Huan Zhang Gui-Ling Zhou Ruo-Yao Ni Yan-Ni Zhao Qi-Lian Qin Zhen Zou State Key Laboratory of Integrated Management of Pest Insects and Rodents Institute of Zoology Chinese Academy of Sciences Beijing China College of Life Sciences Hebei University Baoding Hebei China University of Chinese Academy of Sciences Beijing China

Insects have a large family of C-type lectins involved in cell adhesion, pathogen recognition and activation of immune responses. In this study, 32 transcripts encoding C-type lectin domain proteins (CTLDPs) were identified from the Thitarodes xiaojinensis transcriptome. According to their domain structures, six CTLDPs with one carbohydraterecognition domain (CRD) were classified into the CTL-S subfamily. The other 23 CTLDPs with two CRDs were grouped into the immulectin (IML) subfamily. The remaining three with extra regulatory domains were sorted into the CTL-X subfamily. Phylogenetic analysis showed that CTL-S and CTL-X members from different insects could form orthologous groups. In contrast, no T. xiaojinensis IML orthologues were found in other insects. Remarkable lineage-specific expansion in this subfamily was observed reflecting that these CTLDPs, as important receptors, have evolved diversified members in response to a variety of microbes. Prediction of binding ligands revealed that T. xiaojinensis, a coldadapted species, conserved the ability of CRDs to combine with Ca^2+ to keep its receptors from freezing. Comparative analysis of induction of CTLDP genes after different immune challenges indicated that IMLs might play critical roles in immune defenses. This study examined T. xiaojinensis CTLDPs and provides a basis for further studies of their characteristics.

关键词： binding ligand carbohydrate-recognition domain C-type lectin domain protein expression profiling phylogenetic feature Thitarodes xiaojinensis

Characterization of a new lectin involved in the protoplast regeneration of Bryopsis hypnoides

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Chinese Journal of Oceanology and Limnology 2009年第3期27卷 502-512页

作者：牛建峰王广策吕芳周百成彭光 Key Laboratory of Experimental Marine Biology Institute of OceanologyChinese Academy of Sciences College of Marine Science and Engineering Tianjin University of Science and Technology The Graduate School of Chinese Academy of Sciences

A group of coenocytic marine algae differs from higher plants,whose totipotency depends on an intact cell(or protoplast).Instead,this alga is able to aggregate its extruded protoplasm in sea water and generate new mature *** is thought that lectins play a key role in the aggregation *** purified a lectin associated with the aggregation of cell organelles in Bryopsis *** lectin was ca.27 kDa with a pI between pH 5 and pH *** absence of carbohydrate suggested that the lectin was not a *** hemagglutinating activity(HA) of the lectin was not dependent on the presence of divalent cations and was inhibited by N-Acetylgalactosamine,N-Acetylglucosamine,and the glycoprotein bovine submaxillary *** lectin preferentially agglutinated Gram-negative *** HA of this lectin was stable between pH 4 to pH *** organelles outside the cytoplasm were agglutinated by the addition of lectin solution(0.5 mg ml-1).Our results suggest that the regeneration of *** is mediated by this *** also demonstrated that the formation of cell organelle aggregates was inhibited by nigericin in natural seawater(pH 8.0).Given that nigericin dissipates proton gradients across the membrane,we hypothesize that the aggregation of cell organelles was proton-gradient dependent.

关键词： Bryopsis hypnoides lectin hemagglutinating cell organelle aggregation regeneration

A novel lectin from mushroom Phellodon melaleucus displays hemagglutination activity,and antitumor activity in a B16 melanoma mouse model

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Food Science and Human Wellness 2023年第5期12卷 1885-1892页

作者： Yuanhui Li Peng Wang Zejun Zhang Qinghong Liu Department of Vegetables College of HorticultureChina Agricultural UniversityBeijing 100193China College of Food Science&Nutritional Engineering China Agricultural UniversityBeijing 100193China

A novel lectin(termed PML)was purified from fruiting bodies of the edible mushroom Phellodon melaleucus(division Basidiomycota)by ion exchange,hydrophobic interaction,and gel filtration chromatographies,with overall titer recovery~60%and 20-fold *** displayed hemagglutination activity 13319 units/mg toward rabbit ***-PAGE and gel filtration analyses revealed that PML is a homodimeric lectin with a molecular weight of 28.8 *** hemagglutination activity was not inhibited by various simple sugars or their derivatives,but was enhanced by cations Ca^(2+),Mg^(2+),Zn^(2+),and Cu^(2+).The activity was stable in pH range 6–9 and in the temperature range 20–60°*** dichroism(CD)spectroscopic analysis showed that PML was composed primarily ofβ-sheets with lowα-helix *** a B16 melanoma mouse model,PML treatment significantly inhibited tumor growth,and increased cytokine IL-10 *** findings suggest that PML is a potential anticancer therapeutic agent.

关键词： Phellodon melaleucus lectin Hemagglutination activity Cytokine IL-10 Antitumor activity

VARIATIONS OF lectin-BINDING GLYCOPROTEINS IN EARLY PREGNANT RABBIT EMBRYO

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Science China Chemistry 1992年第5期35卷 577-584页

作者：陈素珍曹咏清 Department of Endocrinology and Laboratory of Reproductive Biology Institute of Zoology Academia Sinica Beijing 100080 PRC

In the present study, the quantitative and qualitative changes of three kinds of lectinbinding glycoproteins of early pregnant rabbit embryos (D4—D12) were analyzed. The technique of Western blot, as well as video densitometer scanning and its analytic software was used for analysis. Results found that there were five specific lectin-binding glycoproteins in Day 4 to Day 6 blastocyst fluids: one ConA-binding glycoprotein about 70 kD, two WGA-binding glycoproteins respectively about 42 kD and 25kD and two PNA-binding glycoprotein respectively about 180kD and 75kD. They disappeared immediately after implan tation, It is demonstrated that there are stage-specific glycoproteins in rabbit blastocyst fluid which might be relevant to the recognition of pregnancy and implantation.

关键词： blastocyst fluid lectin glycoprotein Western blot implantation.