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Acta Pharmacologica Sinica 2005年第12期26卷 1485-1491页

作者： chuan WANG Xiao-na DU Qing-zhong JIA Hai-lin ZHANG~2 Department of Pharmacology Hebei Medical University,Shijiazhuang 050017,china Department of Pharmacology Hebei Medical University Shijiazhuang China

Aim:To investigate the effects of the pleckstrin homology(PH)domain of phos-pholipase cδ1(PLcδ1PH)on inhibition of phospholipase c(PLc)-mediated hy-drolysis of phosphatidylinositol 4,5-bisphosphate[PtdIns(4,5)P2]by ***:A fusion construct of green fluorescent protein(GFP)and PLcδ1PH(PLcδ1PH-GFP),which is known to bind PtdIns(4,5)P2specifically,together withlaser-scanning confocal microscopy,was used to trace PtdIns(4,5)***:Stimulation of the type 1 muscarinic receptor and the bradykinin 2 recep-tor induced a reversible PLcδ1PH-GFP translocation from the membrane to thecytosol in cOS-7 *** inhibitor U73122 blocked the ***,a known PtdIns kinase inhibitor,did not affect the translocationinduced by Ach,but blocked recovery after translocation,indicating thatPtdIns(4,5)P2hydrolysis occurs through receptor-mediated PLc ***,a commonly used phospholipase c blocker,failed to block the recep-tor-induced PLcδ1PH-GFP translocation,indicating that neomycin is unable toblock PLc-mediated Ptdlns(4,5)Pe ***,in the absence of PLcδ1PH-GFP expression,neomycin abolished the receptor-induced hydrolysis ofPtdIns(4,5)P2 by ***:Although PLcδ1PH and neomycin bind toPtdIns(4,5)P2 in a similar way,they have distinct effects on receptor-mediatedactivation of PLc and PtdIns(4,5)P2 hydrolysis.

关键词： phosphatidylinositol 4,5-bisphosphate neomycin phospholipase c pleckstrin homology domains competitive binding acetylcholine green fluorescent proteins

Signaling mechanisms mediated by G-protein coupled receptors in human platelets

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Acta Pharmacologica Sinica 2004年第7期25卷 41-46页

作者： Sheikh Arshad SAEED,Huma RASHEED,Faisal A Wahed FEcTO,Mohammad Ilyas AcHAKZAI,Rahmat ALI,John Dennis cONNOR,Anwar-ul-Hassan GILANI Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan Department of Biological and Biomedical Sciences TheAga Khan University Karachi-74800 Pakistan

AIM: The present study deals with the investigation of mechanisms involved in the synergistic interaction between epinephrine and arachidonic acid (AA). METHODS: Venous blood was taken from healthy human volunteers reported to be free of medications for one week. Platelet aggregation was monitored at 37oc using Dual-channel Lumi-aggregometer. The resulting aggregation was recorded for 5 min by the measurement of light transmission as a function of time. RESULTS: The data show that a synergism in platelet aggregation mediated by subthreshold concentrations of epinephrine (1 μmol/L) and AA (0.2 μmol/L) was inhibited by the α2-receptor antagonist (yohimbine, Ic50=0.6 μmol/L) and an inhibitor of AA-cyclooxygenase (cOX), indomethacin (Ic50=0.25 μmol/L). In examining receptor influence on intraplatelet signalling pathways, it was found that the synergistic effect was inhibited by calcium channel blockers, verapamil (Ic50=0.4 μmol/L) and diltiazem (Ic50=2.5 μmol/L), as well as by low concentrations of inhibitors of phospholipase c (PLc) (U73122; Ic50=0.2 μmol/L) and mitogens activated protein kinase (MAPK) (PD 98059; Ic50=3.8 μmol/L). Herbimycin A, a specific inhibitor of tyrosine light chain kinase (TLcK), showed inhibition at Ic50 value of 15 μmol/L, whereas chelerythrine, a protein kinase c (PKc) inhibitor, had no effect up to 20 μmol/L. cONcLUSION: These data suggest that synergism between epinephrine and AA in platelet aggregation is triggered through receptors coupled to G-protein, which in turn, activate PLc, cOX, and MAP kinase-signaling pathways.

关键词： platelet aggregation epinephrine arachidonic acid cyclooxygenase phospholipase c calcium channel blockers nitric oxide MAP kinase

Phosphoinositide pathway and the signal transduction network in neural development

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Neuroscience Bulletin 2012年第6期28卷 789-800页

作者： Vincenza Rita Lo Vasco Department Organi di Senso Policlinico Umberto IFaculty of MedicineSapienza University of Romeviale del Policlinico 33Rome 00185Italy

The development of the nervous system is under the strict control of a number of signal transduction pathways, often interconnected. Among them, the phosphoinositide （PI） pathway and the related phospholipase c （PI-PLc） family of enzymes have been attracting much attention. Besides their well-known role in the regulation of intracellular calcium levels, PI-PLc enzymes interact with a number of molecules belonging to further signal transduction pathways, contributing to a specific and complex network in the developing nervous system. In this review, the connections of PI signalling with further transduction pathways acting during neural development are discussed, with special regard to the role of the PI-PLc family of enzymes.

关键词： phospholipase c cNS development calcium release

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Regulation of TRP-like muscarinic cation current in gastrointestinal smooth muscle with special reference to PLc/InsP3/ca2+ system

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Acta Pharmacologica Sinica 2006年第7期27卷 833-842页

作者： Alexander V ZHOLOS Department of Physiology Queen’s University Belfast UK

Acetylcholine,the main enteric excitatory neuromuscular transmitter,evokes mem-brane depolarization and contraction of gastrointestinal smooth muscle cells byactivating G protein-coupled muscarinic *** the cholinergic exci-tation is generally underlined by the multiplicity of ion channel effects,the pri-mary event appears to be the opening of cation-selective channels;among themthe 60 pS channel has been recently identified as the main target for the acetylcho-line action in gastrointestinal *** evoked cation current,termed mIcAT,causes either an oscillatory or a more sustained membrane depolarization response,which in turn leads to increases of the open probability of voltage-gated ca2+channels,thus providing ca2+entry in parallel with ca2+release for intracellularca2+concentration rise and *** recent years there have been severalsignificant developments in our understanding of the signaling processes under-lying *** have revealed important synergistic interactionsbetween M2and M3receptor subtypes,single channel mechanisms,and the in-volvement of TRPc-encoded proteins as essential components of native musca-rinic cation *** review summarizes these recent findings and in par-ticular discusses the roles of the phospholipase c/InsP3/intracellular ca2+releasesystem in the mIcATphysiological regulation.

关键词： smooth muscle muscarinic receptor G protein phospholipase c InsP3 calcium TRP cation channel

An Extracellular Oligopeptide Permease May Be a Potential Virulence Factor of Vibrio harveyi

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Journal of Ocean University of china 2011年第4期10卷 343-350页

作者： HE Qingfang cHEN Jixiang LI caifeng Department of Marine Biology College of Marine Life SciencesOcean University of ChinaQingdao 266003P.R.China Oil Production Technology Research Institute Shengli Oilfield Company of SINOPECDongying 257000P.R.China

An oligopeptide permease A(OppA)was purified from the extracellular product of Vibrio harveyi *** molecular weight of the purified protein was estimated to be 58 kDa on *** purified protein showed phospholipase c activity at the optimal values of temperature 50℃ and pH *** enzymatic activity decreased when the temperature increased to 40℃.The N-terminal sequence of the purified protein was determined as ADVPAGTKLA,which is similar to that of *** OppA pre-cursor gene was cloned from the genome of *** *** gene consisted of 1665 base pairs and encoded a 554 amino acid polypeptide,which showed a high similarity to those of the OppAs of *** and other Vibrio *** gene was subcloned into pET-28a(+)and expressed in Escherichia *** expressed recombinant protein was purified by Ni-NTA metal affinity *** expressed recombinant protein showed a 58 kDa band on SDS-PAGE and exhibited phospholipase c activity with the optima of temperature 50℃ and pH *** purified protein was toxic to the flounder gill *** OppA mutant of *** SF-1 was constructed by homologous *** mutant strain was less virulent when it was intraperitoneally inoculated to zebra fish,with the LD50 of 5.46×105 cFU fish-1,compared to 3.11×104 cFU fish-1 of the wild-type strain,which indicated that the OppA might play an important role in the pathogenicity of ***.

关键词： Vibrio harveyi oligopeptide permease phospholipase c pathogenicity

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Preparation of Monoclonal Antibody Against clostridium perfringensα-toxin and Screening and Identification of Phage Display Technology

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Journal of Northeast Agricultural University(English Edition) 2020年第3期27卷 76-86页

作者： Wang Hong-wei Liu Hai-xin Xie Zhen-zhen Zhang Yue Zhong Ming Li Guang-xing Key Laboratory for Laboratory Animals and comparative Medicine of Heilongjiang Province College of Veterinary MedicineNortheast Agricultural UniversityHarbin 150030China

clostridium perfringens phospholipase c(plc),also calledα-toxin,is encoded by the plc gene clostridium *** production ofα-toxin can lead to the occurrence of gas *** is considered as one of the best solutions against clostridium *** this study,an anti-cpα-toxin monoclonal antibody(mAbs)A10E5 was successfully prepared,which had better biological ***,the phage random 12-peptide library was used to screen mAb A10E5 *** four rounds of screening,three peptides with high affinity to the anti-α-toxin mAbs were *** 12-peptide peptide Q and peptide E with higher inhibition rate were obtained by indirect *** polypeptides of 500μg·mL^(-1)synthesized in vitro were mixed with30μg·mL^(-1)α-toxin at a concentration to treat Hela *** viability was determined by MTT *** results showed that both of the peptides significantly increased the survival rate of Hela cells compared with theα-toxin group,and the effect of peptide Q was more *** chickens were immunized with phages expressing two different affinity polypeptides and then *** results of chicken weight change,intestinal lesion score,bacterial count,and antibody titer in peripheral blood showed that the two phages expressing the polypeptides had a certain protective effect on the chickens compared with the PBS group,and peptide Q had better protection *** conclusion,the high affinity peptide with mAb A10E5 was screened in this study,and the protective effect of the plc polypeptide vaccine was verified by in vivo and in vitro experiments,which was of great significance for the comprehensive prevention and treatment of the disease.

关键词： clostridium perfringens phospholipase c phage peptide

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The Difference in calcium Levels in Aspergillus nidulans Grown on Glucose or Pectin

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Advances in Microbiology 2012年第2期2卷 117-121页

作者： Janice Aparecida Rafael Suraia Said Departamento de ciencias Farmaceuticas Faculdade de Ciencias Farmaceuticas de Ribeirao PretoUniversidade de Sao PauloSao PauloBrazil

Understanding the growth regulatory mechanisms in filamentous fungi is very important for the production of medicines for antifungal therapies. It is well established that ca2+ gradient is essential for hyphal growth and that one mechanism responsible for the ca2+ cellular concentration starts with the hydrolysis of phosphatidylinositol 4,5-bisphosphate (PIP2) by receptor-regulated forms of phosphoinositide-specific phospholipase c (PI-PLc). In the present study the levels of calcium in Aspergillus nidulans wild type (A26) and plcA-deficient mutant (AP27) growing in a carbon source readily assimilated, as glucose or pectin a non-readily assimilated carbon source was investigated. Intracellular calcium levels in A26 were higher in the presence of glucose than in pectin, but lower in AP27 independently of the carbon source and in AP27 the vesicular calcium distribution occurred mainly at the apex of the hyphae. Delay in nuclear division was also observed if A26 and AP27 were grown in pectin presence when compared with growth in glucose. For the first time, it is demonstrated that the levels of intracellular ca2+ were higher when A. nidulans was growing in glucose than in a non readily assimilated carbon source as pectin. Further, it also showed that the plcA gene, although not essential, may be responsible for high-molecular weight carbon source recongnation, for the intracellular ca2+ levels maintenance and consequently by the nuclear division in A. nidulans.

关键词： Aspergillus nidulans ca^(2+) Signaling phospholipase c

Function Analysis of AtPLc6 Regulates Pollen Tube Tip Growth in Arabidopsis thaliana

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Function Analysis of AtPLC6 Regulates Pollen Tube Tip Growth...

Ghrelin enhances the excitability of nigral dopaminergic neurons by activating its receptors and inhibiting potassium channels via PLc/PKc pathway

中国植物生理学会第十次会员代表大会暨全国学术年会

作者： Juan Du, Min-juan Lan, Li-min Lv, Yan-yun Pan* college of Life Sciences, Hebei Agricultural University, Baoding 071000, china

Pollen development involves complex developmental control of gene expression by the haploid genome[1]. Phosphoinositide-specific phospholipase c is a key role in the phosphoinositide signaling transduction

关键词： Arabidopsis thal iana phospholipase c pollen germination

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Mechanical Strain regulates osteoblast proliferation through ca2+-caMK-cREB Signal Pathway

中国生理学会第23届全国会员代表大会暨生理学学术大会

作者： Li-Min SHI,Xi-Xun DU,Zhi-Qiang QU,Ze-Gang MA,Yu ZHOU,Hong JIANG,Jun-Xia XIE~* Department of Physiology,Shandong Provincial Key Laboratory of Pathogenesis and Prevention of Neurological Disorders and State Key Disciplines:Physiology,Medical college of Qingdao University,Qingdao,266071

＜正＞Excitability of dopaminergic neurons in substantia nigra pars compacta（SNc） is associated with neuroprotection and motor *** previously found that ghrelin protected dopaminergic neurons against neurotoxicity by inhibiting apoptosis and attenuating the oxidative ***,its physiological mechanism remains *** this study,using rat brain slices,we observed that ghrelin at physiological concentrations not only increased the spontaneous firing rate of nigral dopaminergic neurons,but also altered their firing from regular pacemaker-like to burst *** evidence suggests that ghrelin enhances the excitability of nigral dopaminergic neurons by activating its receptors（GHS-R1a）/PLc/PKc pathway since D-[Lys3]-GHRP-6,a selective antagonist for GHS-R1a,PLc inhibitor U-73122 or PKc inhibitor GF109203X fully abolished the ghrelin effect;potassium channels are considered to be a final effector of this pathway because tetraethylammol/Lonium chloride totally blocked ghrelin-induced excitability *** significantly altered the mIPScs,indicating that ghrelin also regulates inhibitory synaptic *** order to reveal any role of ghrelin in motor coordination,we examined the effect of ghrelin on animal *** microinjection of ghrelin into the SNc caused contralateral dystonic posturing and attenuated catalepsy elicited by systemic haloperidol administration in vivo. These motor effects may be due to increase of dopamine（DA） release from SNc because intracerebroventricular administration of ghrelin caused increase of DA turnover rate in the ***,the results suggest that ghrelin,as a physiological mediator capable of regulating neuronal excitability,participates in the motor function and neuroprotection in SNc.

关键词： ghrelin dopaminergic neurons cell excitability GHS-R1a phospholipase c mIPScs

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Mechanical Strain regulates osteoblast proliferation through...