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Characterization of Angiotensin-? Converting Enzyme Inhibiting Peptide from venerupis philippinarum with Nano-Liquid Chromatography in Combination with Orbitrap Mass Spectrum Detection and Molecular Docking

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Journal of Ocean University of China 2017年第3期16卷 473-478页

作者： SHI Lei WU Tizhi SHENG Naijuan YANG Li WANG Qian LIU Rui WU Hao Jiangsu Key Laboratory of Research and Development in Marine Bio-resource Pharmaceutics Nanjing University of Chinese MedicineNanjing 210023P.R.China Jiangsu Collaborative Innovation Center of Chinese Medicinal Resources Industrialization and National and Local Collaborative Engineering Center of Chinese Medicinal Resources Industrialization and Formulae Innovative MedicineNanjing University of Chinese MedicineNanjing 210023P.R.China

The complexity and diversity of peptide mixture from protein hydrolysates make their characterization difficult. In this study, a method combining nano LC-MS/MS with molecular docking was applied to identifying and characterizing a peptide with angiotensin-? converting enzyme(ACE-I) inhibiting activity from venerupis philippinarum hydrolysate. Firstly, ethanol supernatant of V. philippinarum hydrolysate was separated into active fractions with chromatographic methods such as ion-exchange chromatography and high performance liquid chromatography in combination. Then seven peptides from active fraction were identified according to the searching result of the MS/MS spectra against protein databases. Peptides were synthesized and subjected to ACE-Iinhibition assay. The peptide NTLTLIDTGIGMTK showed the highest potency with an IC_(50) of 5.75 μmol L^(-1). The molecular docking analysis showed that the ACE-I inhibiting peptide NTLTLIDTGIGMTK bond with residues Glu123, Glu403, Arg522, Glu376, Gln281 and Asn285 of ACE-I. Therefore, active peptides could be identified with the present method rather than the traditional purification and identification strategies. It may also be feasible to identify other food-derived peptides which target other enzymes and receptors with the method developed in this study.

关键词： ACE-I inhibitor ACE-I inhibiting peptide characterization nano-LC-MS/MS molecular docking venerupis philippinarum

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Molecular cloning and differential expression patterns of sigma and omega glutathione S-transferases from venerupis philippinarum to heavy metals and benzo[a]pyrene exposure

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Chinese Journal of Oceanology and Limnology 2012年第3期30卷 413-423页

作者：张林宝吴惠丰刘小莉陈磊磊王清赵建民由丽萍 Key Laboratory of Coastal Zone Environmental Processes Yantai Institute of Coastal Zone Research(YIC)Chinese Academy of Sciences(CAS) Shandong Provincial Key Laboratory of Coastal Zone Environmental Processes YICCAS Graduate University of Chinese Academy of Sciences

Glutathione S-transferases (GSTs) are a class of enzymes that facilitate the detoxification of xenobiotics, and also play important roles in antioxidant defense. We identified two glutathione S-transferase isoforms (VpGSTS, sigma GST; VpGSTO, omega GST) from venerupis philippinarum by RACE approaches. The open reading frames of VpGSTS and VpGSTO were of 612 bp and 729 bp, encoding 203 and 242 amino acids with an estimated molecular mass of 22.88 and 27.94 kDa, respectively. The expression profiles of VpGSTS and VpGSTO responded to heavy metals and benzo[a]pyrene (B[a]P) exposure were investigated by quantitative real-time RT-PCR. The expression of VpGSTS and VpGSTO were both rapidly up-regulated, however, they showed differential expression patterns to different toxicants. Cd displayed stronger induction of VpGSTS expression with an approximately 12-fold increase than that of VpGSTO with a maximum 6.4-fold rise. Cu exposure resulted in similar expression patterns for both VpGSTS and VpGSTO. For B[a]P exposure, the maximum induction of VpGSTO was approximately two times higher than that of VpGSTS. Altogether, these findings implied the involvement of VpGSTS and VpGSTO in host antioxidant responses, and highlighted their potential as a biomarker to Cd and B[a]P exposure.

关键词： venerupis philippinarum Glutathione S-transferase mRNA expression heavy metals benzo[a]pyrene biomarker

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The first molluscan TCCP in venerupis philippinarum:molecular cloning and expression analysis

The first molluscan TCCP in Venerupis philippinarum:molecula...

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中国动物学会、中国海洋湖沼学会贝类学会分会第十四次学会研讨会

作者： Chenghua Li~a,Lihua Qiu~b,Aiqin Chen~a,Jianmin Zhao~(a*),Song Qin~a (a.Yantai Institute of Coastal Zone Research,Chinese Academy of Sciences,Yantai,264003, China b.South China Sea Fishery Research Institute,Chinese Academy of Fishery Sciences, Guangzhou,510301,China)

Translationally controlled tumour protein(TCTP)is one of the abundant and ubiquitously expressed proteins in *** the present study,the first moiluscan TCCP of venerupis philippinarum(denoted as VpTCTP)was identified from haemocytes by EST and RACR *** full-length cDNA of VpTCTP consisted of 1148 nucleotides with a canonical polyadenylation signal sequence AATAAA,a polyA tail,and an open-reading frame of 555 bp encoding 184 amino *** deduced amino acid of VpTCTP shared high similarity with the TCTPs from other species,indicating that VpTCTP should be a member of TCTPs family. Several highly conserved motifs including 5' UTR starting with 5'terminal ologopyrimidine (5'TOP),3'UTR rich in AU and AUUT elements were also identified in *** tissue and temporal expression of VpTCTP after Vibrio anguillarum challenge was recorded by quantitative real-time *** transcript could be detected in all examined tissues including hepatopancreas,gill,muscle,heamocyte and foot with the highest expression level in heamocytes and lowest in *** the time-course expression in heamocytes,the relative expression level of VpTCTP mRNA was down-regulated sharply from 6 hr to 12 hr post-infection. Then,the expression level increased and reached to 1.2-fold compared to the control group at 72 *** 96 hr,the expression drop back to the original *** these results indicated that VpTCTP was an acute-phase protein involved in the immune response of ***.

关键词： venerupis philippinarum TCTP Tissues expression Time-course expression

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Two classes of glutathione S-transferase genes with different response profiles to bacterial challenge in venerupis philippinarum

Two classes of glutathione S-transferase genes with differen...

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中国动物学会·中国海洋湖沼学会贝类学分会第九次会员代表大会暨第十五次学术讨论会

作者： Chenghua Li~(a*) Xiurong Su~(a*) Ye Li~(ab),Taiwu Li~(ab) Chongjie Sun~a Tingting Zhou~a a Faculty of Life Science and Biotechnology,Ningbo University,Ningbo b Ningbo City College of Vocational Technology,Ningbo

Glutathione S-transferase(GST) is major cytosolic detoxification enzymes involved in many pathological and physiological *** the present study,two classes of GSTs(VpGST-1 and VpGST-2) were cloned from venerupis philippinarum haemocytes by cDNA library and RACE *** alignments and phylogenetic analysis together supported that they belonged to a new member of sigma and pi classes GSTs protein family,*** expression profiles of these two genes under Vibrio anguillarum challenge were investigated by quantitative *** bacterial challenge could significantly up-regulate the mRNA expression of both VpGST-1 and VpGST-2 with larger amplitude in VpGST-2,and the feedback speed for VpGST-2 was more rapid than that of *** differences in the response to bacterial challenge indicated that they were functional diversity and probably played cooperative roles in mediating the Vibrio challenge in clam.

关键词： venerupis philippinarum Glutathione S-transferase Expression profile Vibrio anguillarum

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Identification and assessment of biochemistry markers in gill and haemocytes of venerupis philippinarum under mercury exposure

Identification and assessment of biochemistry markers in gil...

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“全球变化下的海洋与湖沼生态安全”学术交流会

作者： Xiaocong CHEN Chenghua LI Ran ZHANG Pengjuan ZHANG Yina SHAO School of Marine Sciences Ningbo University

The impact of mercury stress on venerupis philippinarum gill and haemocytes was assessed through investigating the dynamic changes of several biochemistry parameters, including superoxide dismutase（SOD）, catalase（CAT）, glutathione reductase（GR）, acid phosphatase（ACP）, alkaline phosphatase（ALP）, total glutathione content, as well as malondialdehyde（MDA） content, under different Hg2+ acute exposure in the present study. Results showed that SOD activities and GSH contents in the gill displayed depressed expression. The correlation analysis indicated significant positive correlations of the two parameters（0.833, P ＜ 0.01）. GR showed elevated activities in all examined exposure doses. While the activities of CAT and ALP were only induced at higher Hg2+ concentrations with significant positive correlations（0.917, P ＜ 0.01）, and MDA showed decreased contents in middle exposure doses. In term of haemocytes, Hg exposure had an induced effect on the activities of CAT, GR, ACP and ALP, and MDA content increased with increasing concentration of Hg at 24 h. All these parameters had significant positive correlations（P ＜ 0.01）. No obvious dose-dependent or time-dependent expression profiles were detected in other indexes in both tissues. All these results showed that acute Hg exposure led to harmful effects on clams immune system, which are likely linked to Hg induced oxidative stress. CAT, ACP, and MDA in gill have great potential as biomarkers of Hg concentration determination in the aquatic environment for the studied species.

关键词： venerupis philippinarum mercury gill haemocytes biomarker

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胶州湾两种习见帘蛤(venerupis semidecussata和venerupis philippinarum)的生殖周期

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山东海洋学院学报 1962年 43-64页

作者：李嘉泳邹仁林王秋杨德渐

1.为进行生殖周期研究的需要,本文首先对胶州湾两种习见帘蛤动物的种名重新进行了鉴定,发现其中原来定名为杂色蛤venerupis variegata的一种应是半叉蛤***。其另一种仍为蛤子***。 2.本文以组织学和细胞学方法为主,研究了胶州湾两种习... 详细信息

1.为进行生殖周期研究的需要,本文首先对胶州湾两种习见帘蛤动物的种名重新进行了鉴定,发现其中原来定名为杂色蛤venerupis variegata的一种应是半叉蛤***。其另一种仍为蛤子***。 2.本文以组织学和细胞学方法为主,研究了胶州湾两种习见帘蛤动物的性腺发育周期,并根据性细胞的发育过程,把它们的性腺发育周期分为:A(增殖期)、B(生长期)、C(成熟期)、D(生殖期)、E(后生殖期)等五个时期。 3.半叉蛤和蛤子雌性腺发育周期的形态学过程彼此完全没有什么差别。此外,两者雄性腺体的发育也有同样表现。这些都是我们进一步考虑它们两者分类关系的依据。 4.结合性腺组织学检查、室内自然产卵观察,人工授精实验、性腺发育指政(平均体重与平均壳长比值)的测定和性腺解剖观察等方法所获得的结果,可以肯定胶州湾半叉蛤和蛤子的生殖周期是一年一次。其生殖季节是从9月上旬到10月中旬,共约一个半月左右的时间。 5.在雄性个体产卵以后,泡囊内仍然残留一部分卵子。鉴于大量吞噬细胞的侵入,这部分卵子继续成熟和产出的可能是不大的。 6.雄性排精活动可能较产卵时间要长一些。甚至到12月底前后,仍有个别个体的泡囊内残留极少精子。但新精子的产生必须到4月初旬左右才能开始。 7.在卵母细胞早期发育过

关键词：帘蛤性腺发育蛤子生殖周期 venerupis semidecussata venerupis philippinarum 胶州湾胶澳两种

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Glutathione S-transferase(GST) gene expression profiles in two marine bivalves exposed to BDE-47 and their potential molecular mechanisms

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Chinese Journal of Oceanology and Limnology 2015年第3期33卷 705-713页

作者：李斐吴惠丰王清李雪花赵建民 Key Laboratory of Coastal Zone Environmental Processes and Ecological Remediation Yantai Institute of Coastal Zone Research (YIC )Chinese Academy of Sciences Shandong Provincial Key Laboratory of Coastal Zone Environmental Processes Key Laboratory of Coastal Zone Environmental Processes and Ecological Remediation Yantai Institute of Coastal Zone Research (YIC) Chinese Academy of Sciences Key Laboratory of Industrial Ecology and Environmental Engineering (MOE) School of Environmental Science and TechnologyDalian University of Technology

Glutathione S-transferases （GSTs） are phase II enzymes that facilitate the detoxification of xenobioties and play important roles in antioxidant defense. We investigated the expression patterns of seven venerupis philippinarum GSTs （VpGSTs） and four Mytilus galloprovincialis GSTs （MgGSTs） following exposure to BDE-47. Differential expressions of the seven VpGSTs and four MgGSTs transcripts were observed, with differences between the hepatopancreas and gills. Among these GSTs, the sigma classes （VpGSTS1, VpGSTS2, VpGSTS3, MgGST1, and MgGST3） were highly expressed in response to BDE-47 exposure, demonstrating their potential as molecular biomarkers for environmental biomonitoring studies. We obtained the three-dimensional crystal structures of VpGSTs and MgGSTs by homologous modeling. A model to elucidate the binding interactions between the ligands and receptors was defined by molecular docking, Hydrophobic and n were the most often observed interactions between BDE-47 and the GSTs.

关键词： glutathione S-transferase (GST) venerupis philippinarum Mytilus galloprovincialis 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) molecular docking biomarker

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