哈维氏弧菌磷脂酶D对不同磷脂单分子层的吸附动力学研究
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摘要
基于单分子层技术研究了哈维氏弧菌来源磷脂酶D(Vh PLD)对不同磷脂单分子层的吸附动力学。探究初始表面压力条件对VhPLD吸附不同磷脂单分子层(磷脂酰胆碱(PC)、磷脂酰乙醇胺(PE)、磷脂酰丝氨酸(PS)、磷脂酰甘油(PG)和磷脂酰肌醇(PI))吸附动力学参数(k_a、k_d、K_(Ads))的影响规律。结果表明:VhPLD对磷脂单分子层的吸附动力学参数与磷脂单分子层初始表面压力密切相关;在15 m N/m条件下,VhPLD对不同磷脂单分子层吸附偏好性顺序为PC> PG> PS> PE=PI;在20 m N/m条件下,VhPLD对不同磷脂单分子层吸附偏好性顺序为PG> PI> PC> PS> PE;在25 m N/m条件下,VhPLD对不同磷脂单分子层吸附偏好性顺序则转变为PC> PS> PI> PE=PG。
The adsorption kinetics of Vibrio harveyi phospholipid D (Vh PLD) to different phospholipid monolayers were investigated using monolayer technology. The adsorption kinetic parameters of Vh PLD under different initial surface pressures to different phospholipid monolayers (Phosphatidylcholine (PC),Phosphatidylethanolamine (PE),Phosphatidylserine (PS),Phosphatidylglycerol (PG) and Phosphatidylinositol (PI)) were investigated,respectively. The results indicated that VhPLD adsorption kinetics parameters to phospholipid monolayer were closely related to initial surface pressure. Under the condition of 15 m N/m,the affinity order of VhPLD to various phospholipid monolayers was PC > PG > PS > PE =PI. Under the condition of 20 m N/m,the affinity order of VhPLD to various phospholipid monolayers was PG > PI > PC > PS > PE. Under the condition of 25 m N/m,the affinity order of VhPLD to various phospholipid monolayers changed into PC > PS > PI > PE = PG.
The adsorption kinetics of Vibrio harveyi phospholipid D (Vh PLD) to different phospholipid monolayers were investigated using monolayer technology. The adsorption kinetic parameters of Vh PLD under different initial surface pressures to different phospholipid monolayers (Phosphatidylcholine (PC),Phosphatidylethanolamine (PE),Phosphatidylserine (PS),Phosphatidylglycerol (PG) and Phosphatidylinositol (PI)) were investigated,respectively. The results indicated that VhPLD adsorption kinetics parameters to phospholipid monolayer were closely related to initial surface pressure. Under the condition of 15 m N/m,the affinity order of VhPLD to various phospholipid monolayers was PC > PG > PS > PE =PI. Under the condition of 20 m N/m,the affinity order of VhPLD to various phospholipid monolayers was PG > PI > PC > PS > PE. Under the condition of 25 m N/m,the affinity order of VhPLD to various phospholipid monolayers changed into PC > PS > PI > PE = PG.
引文
[1]杨学利,马沁沁,袁向华,等.磷脂酶D及其催化机制的研究进展[J].中国油脂,2016,41(3):80-84.
[2]LERCHNER A,MANSFELD J,SCHFFNER I,et al.Two highly homologous phospholipase D isoenzymes from Papaver somniferum L.with different transphosphatidylation potential[J].Biochim Biophys Acta,2005,1737(2/3):94-101.
[3]DAMNJANOVIC J,IWASAKI Y.Phospholipase D as a catalyst:application in phospholipid synthesis,molecular structure and protein engineering[J].J Biosci Bioeng,2013,116(3):271-280.
[4]MOMSEN W E,BROCKMA H L.The adsorption to and hydrolysis of 1,3-didecanoyl glycerol monolayers by pancreatic lipase.Effects of substrate packing density[J].JBiol Chem,1981,256(13):6913-6916.
[5]KUPPE K,KERTH A,BLUME A,et al.Calcium-induced membrane microdomains trigger plant phospholipase D activity[J].Chem Bio Chem,2008,9(17):2853-2859.
[6]BREZESINSKI G,MOHWALD H.Langmuir monolayers to study interactions at model membrane surfaces[J].Adv Colloid Interface Sci,2003(100/102):563-584.
[7]BROCKMA H.Lipid monolayers:why use half a membrane to characterize protein-membrane interactions?[J].Curr Opin Struct Biol,1999,9(4):438-443.
[8]BOUCHER J,TRUDEL E,METHOT M,et al.Organization,structure and activity of proteins in monolayers[J].Colloids Surf B Biointerfaces,2007,58(2):73-90.
[9]MAGET-DANA R.The monolayer technique:a potent tool for studying the interfacial properties of antimicrobial and membrane-lytic peptides and their interactions with lipid membranes[J].Biochim Biophys Acta,1999,1462(1/2):109-140.
[10]BNAROUCHE A,POINT V,PARSIEGLA G,et al.New insights into the p H-dependent interfacial adsorption of dog gastric lipase using the monolayer technique[J].Colloids Surf B Biointerfaces,2013,111:306-312.
[11]BARQUE J P,DERVICHIAN D G.Enzyme-substrate interaction in lipid monolayers.II.Binding and activity of lipase in relation to enzyme and substrate concentration and to other factors[J].J Lipid Res,1979,20(4):447-455.
[12]PHILLIPS M C,SPARKS C E.Properties of apolipoprotens at the air-water interface[J].Ann N Y Acad Sci,1980,348(1):122-137.
[13]SAYARI A,MOSBAH H,VERGER R,et al.The N-terminal His-tag affects the enantioselectivity of staphylococcal lipases:a monolayer study[J].J Colloid Interface Sci,2007,313(1):261-267.
[14]BOISSELIER,DEMERS,CANTIN L,et al.How to gather useful and valuable information from protein binding measurements using Langmuir lipid monolayers[J].Adv Colloid Interface Sci,2017,243:60-76.
[2]LERCHNER A,MANSFELD J,SCHFFNER I,et al.Two highly homologous phospholipase D isoenzymes from Papaver somniferum L.with different transphosphatidylation potential[J].Biochim Biophys Acta,2005,1737(2/3):94-101.
[3]DAMNJANOVIC J,IWASAKI Y.Phospholipase D as a catalyst:application in phospholipid synthesis,molecular structure and protein engineering[J].J Biosci Bioeng,2013,116(3):271-280.
[4]MOMSEN W E,BROCKMA H L.The adsorption to and hydrolysis of 1,3-didecanoyl glycerol monolayers by pancreatic lipase.Effects of substrate packing density[J].JBiol Chem,1981,256(13):6913-6916.
[5]KUPPE K,KERTH A,BLUME A,et al.Calcium-induced membrane microdomains trigger plant phospholipase D activity[J].Chem Bio Chem,2008,9(17):2853-2859.
[6]BREZESINSKI G,MOHWALD H.Langmuir monolayers to study interactions at model membrane surfaces[J].Adv Colloid Interface Sci,2003(100/102):563-584.
[7]BROCKMA H.Lipid monolayers:why use half a membrane to characterize protein-membrane interactions?[J].Curr Opin Struct Biol,1999,9(4):438-443.
[8]BOUCHER J,TRUDEL E,METHOT M,et al.Organization,structure and activity of proteins in monolayers[J].Colloids Surf B Biointerfaces,2007,58(2):73-90.
[9]MAGET-DANA R.The monolayer technique:a potent tool for studying the interfacial properties of antimicrobial and membrane-lytic peptides and their interactions with lipid membranes[J].Biochim Biophys Acta,1999,1462(1/2):109-140.
[10]BNAROUCHE A,POINT V,PARSIEGLA G,et al.New insights into the p H-dependent interfacial adsorption of dog gastric lipase using the monolayer technique[J].Colloids Surf B Biointerfaces,2013,111:306-312.
[11]BARQUE J P,DERVICHIAN D G.Enzyme-substrate interaction in lipid monolayers.II.Binding and activity of lipase in relation to enzyme and substrate concentration and to other factors[J].J Lipid Res,1979,20(4):447-455.
[12]PHILLIPS M C,SPARKS C E.Properties of apolipoprotens at the air-water interface[J].Ann N Y Acad Sci,1980,348(1):122-137.
[13]SAYARI A,MOSBAH H,VERGER R,et al.The N-terminal His-tag affects the enantioselectivity of staphylococcal lipases:a monolayer study[J].J Colloid Interface Sci,2007,313(1):261-267.
[14]BOISSELIER,DEMERS,CANTIN L,et al.How to gather useful and valuable information from protein binding measurements using Langmuir lipid monolayers[J].Adv Colloid Interface Sci,2017,243:60-76.