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P-D1-02 Fluorescence assay for the specific binding of C-reactive protein with model receptors
Dl Structure of complex assemblies P-Dl-02 FLU$Ht%KJBm,ASSAY FOR Tt$lkS@@CiFiC BIb@~G OF C-B&ACTIVE FR0TEIrir MODEL RECEPTORS ZHEN43LlU. SEN-FANGSUI Dept.Biol.Sci.t Biol..TsinghuaUniv..Beijing(CN).
P-D141
SEN-FANGSUI.ZHENG LlU. CAIDE XLQO Dept.Biol.sci.& Biol..TsingbuaUniv.,Beijing(CN). PurpasetC-reactiveprotein(CRP)is one of the mostchrrscberistic acutephaseproteins.We report herethe two dimensionalcrystalliition of CRPand the stntctm analysisby electroncrystallography. Method: Baseupon the interactions between water solubleproteinand Iipid monolayersCRP2D crystalswereorgan&d. For CRPspecificbinding,a long-armspacercontainingPC wassynthesized.In the cllseof non-specificadsorptionthe electrostatic interactionwasfmely controlled. RemIts: Two typesofcrystaJlinearraysof CRP were obtainedwith two methods.Electroncrystallographicanalysisof the negatively-stainedspecimens showedthat the unit cellparametersof the CRP2D crystalsformedby specificbinding werea=8l& b-78,&, y=61.6S0, and that formed by non-specific adsorptionwerea=744 b=67A,~95.5 O.A resolution of the projectionmapsat 2ti wasobtained. Conclusions:Both projectionmapsof the two types 2D crystalsexhibitedthat only the subunitsof the CRPwerepackedin 2D arraysand the orientations of the subunitson differentmonolayerswere not same.By comparingthe two projectionmaps,a /preliminaryshapeof the CRPsubunithasbeen derived.A modelof the pentamericstructureof the oligomtic CRPwassuggested.
Purpose:C-reactiveprotein(CRP)is one of the mostcharacteristicacutephaseproteinsin human and in manyother animals.previousstudyshowed that the intactlipid membranepreventsthe specific bindingbetweenCBPand PCgroups.Using two syntheticamphiphilicmoleculeswith a long-arm spacerinsertedbetweenthe PChead group andthe hydrophobictailsasthe membranebound model receptors,we studiedthe primaryfactor effected CRPbinding. Method: A fluorescenceassaywas developedfor the specificbindingof CRPwith its receptors,which was uponthe fluorescencequenchingof tyrosines aroundthe binding siteof CRP. Resulti The fluorescencespectrashowedthat the fluorescenceintensityof CRPwas remarkably decreasedafterspecificallybinding with its receptors,phosphorylchome,and with the synthetic amphiphilicmoleculescontainingliposomes. Conclusions:The fluorescenceassayfor CFtP binclmgshowedthat CRPcould specificallybind onto the surfaceof the membranebound model receptorcontainingliposomesand the steric hindranceis oneof the mostimportant factorsthat influencesthis specificbinding.
P-Dl-03 AGGREGATION DUE ‘t’0 THE INTERACTION OF BIOMACRt$Mt#&CULES WITH ORGANIC DYES fN WA’ttR !t&IXJllONS BORISSEVITCH I, TAFJAKM. Inst. de Quhica de SHO carlos,Univ.deSILO Paul0(BR)
P-Dl-04
ONTHESTRUCWREANDFUNCTION OFFISN-TERMINAL; ACOMPUTER MODELINGPREDICTION HWANG MJ, TZOU WS. Inst.of Biomed.sci., Academia Siniq Taipei(TW)
Purpose: Binding of organic dyes with biosysternscan provoke ~ggre@on. This should change physico-chu!uicaIch&&ristics of both partners. Methods: This was demtmstmtd by optical absorption, fluorescence,‘H NMR, &R.a~d r&ance light scaltering speobuscopieson the example of water solublepotphyrins and p&chromophoric cytmine dyes in the pmscne C&DNA and bovine sertmt albumin in aqueousst~lutions. Results: TWO types ‘of aggregation were demonstrated: the dye aggregation on tbe~macromolecule surface and the Womolecule aggregation around the dye. Tbe chamc&Wcs of the aggregates depend on the am&ures of the dye and the macromolecule. The wg@on ohangesthe reaction abilities, pammag&c and spectral propertiesofthedyeandeniergetid~t@necbarkteristics of their singlet and triplet excited states. Conclusions: The aggregation is a common phenomemon at the interaction of biomacromolecules with organic dyes. This should be taken into account at their medical application. Support: CNPq, FlNEP, FAPESP(BR) 139
Purpose: Our understanding of, F&&ted DNA inversion is severelyhandicapped by the missing of a structure for the Fis N-terminal domain (-20 residues) which confers the protein’s stimulation tiction through generally believed, a direct protein-protein interaction with invertase. Methods: A combined sequence/structurehomology analysisagainst protein data bank was used to identify the most probable conformation for the Fis N-terminal. BLAST and TOPITS were the methodsused in the analysis. Results: A loop structure of flap-hinge type for the elusiveFis N-terminal was predicted. The prediction appears to fit mutagenesis experiments-derivedevidenceon this region. Conclusions: A stmctural model for the Fis Nterminal is predicted, making it po%ible, for the first time, for one to model the putative Fisinvettase complex in the continuing kfforts to unveil the molecular mechanism of invertase-, mediatedDNA recombination.
P-D141
SEN-FANGSUI.ZHENG LlU. CAIDE XLQO Dept.Biol.sci.& Biol..TsingbuaUniv.,Beijing(CN). PurpasetC-reactiveprotein(CRP)is one of the mostchrrscberistic acutephaseproteins.We report herethe two dimensionalcrystalliition of CRPand the stntctm analysisby electroncrystallography. Method: Baseupon the interactions between water solubleproteinand Iipid monolayersCRP2D crystalswereorgan&d. For CRPspecificbinding,a long-armspacercontainingPC wassynthesized.In the cllseof non-specificadsorptionthe electrostatic interactionwasfmely controlled. RemIts: Two typesofcrystaJlinearraysof CRP were obtainedwith two methods.Electroncrystallographicanalysisof the negatively-stainedspecimens showedthat the unit cellparametersof the CRP2D crystalsformedby specificbinding werea=8l& b-78,&, y=61.6S0, and that formed by non-specific adsorptionwerea=744 b=67A,~95.5 O.A resolution of the projectionmapsat 2ti wasobtained. Conclusions:Both projectionmapsof the two types 2D crystalsexhibitedthat only the subunitsof the CRPwerepackedin 2D arraysand the orientations of the subunitson differentmonolayerswere not same.By comparingthe two projectionmaps,a /preliminaryshapeof the CRPsubunithasbeen derived.A modelof the pentamericstructureof the oligomtic CRPwassuggested.
Purpose:C-reactiveprotein(CRP)is one of the mostcharacteristicacutephaseproteinsin human and in manyother animals.previousstudyshowed that the intactlipid membranepreventsthe specific bindingbetweenCBPand PCgroups.Using two syntheticamphiphilicmoleculeswith a long-arm spacerinsertedbetweenthe PChead group andthe hydrophobictailsasthe membranebound model receptors,we studiedthe primaryfactor effected CRPbinding. Method: A fluorescenceassaywas developedfor the specificbindingof CRPwith its receptors,which was uponthe fluorescencequenchingof tyrosines aroundthe binding siteof CRP. Resulti The fluorescencespectrashowedthat the fluorescenceintensityof CRPwas remarkably decreasedafterspecificallybinding with its receptors,phosphorylchome,and with the synthetic amphiphilicmoleculescontainingliposomes. Conclusions:The fluorescenceassayfor CFtP binclmgshowedthat CRPcould specificallybind onto the surfaceof the membranebound model receptorcontainingliposomesand the steric hindranceis oneof the mostimportant factorsthat influencesthis specificbinding.
P-Dl-03 AGGREGATION DUE ‘t’0 THE INTERACTION OF BIOMACRt$Mt#&CULES WITH ORGANIC DYES fN WA’ttR !t&IXJllONS BORISSEVITCH I, TAFJAKM. Inst. de Quhica de SHO carlos,Univ.deSILO Paul0(BR)
P-Dl-04
ONTHESTRUCWREANDFUNCTION OFFISN-TERMINAL; ACOMPUTER MODELINGPREDICTION HWANG MJ, TZOU WS. Inst.of Biomed.sci., Academia Siniq Taipei(TW)
Purpose: Binding of organic dyes with biosysternscan provoke ~ggre@on. This should change physico-chu!uicaIch&&ristics of both partners. Methods: This was demtmstmtd by optical absorption, fluorescence,‘H NMR, &R.a~d r&ance light scaltering speobuscopieson the example of water solublepotphyrins and p&chromophoric cytmine dyes in the pmscne C&DNA and bovine sertmt albumin in aqueousst~lutions. Results: TWO types ‘of aggregation were demonstrated: the dye aggregation on tbe~macromolecule surface and the Womolecule aggregation around the dye. Tbe chamc&Wcs of the aggregates depend on the am&ures of the dye and the macromolecule. The wg@on ohangesthe reaction abilities, pammag&c and spectral propertiesofthedyeandeniergetid~t@necbarkteristics of their singlet and triplet excited states. Conclusions: The aggregation is a common phenomemon at the interaction of biomacromolecules with organic dyes. This should be taken into account at their medical application. Support: CNPq, FlNEP, FAPESP(BR) 139
Purpose: Our understanding of, F&&ted DNA inversion is severelyhandicapped by the missing of a structure for the Fis N-terminal domain (-20 residues) which confers the protein’s stimulation tiction through generally believed, a direct protein-protein interaction with invertase. Methods: A combined sequence/structurehomology analysisagainst protein data bank was used to identify the most probable conformation for the Fis N-terminal. BLAST and TOPITS were the methodsused in the analysis. Results: A loop structure of flap-hinge type for the elusiveFis N-terminal was predicted. The prediction appears to fit mutagenesis experiments-derivedevidenceon this region. Conclusions: A stmctural model for the Fis Nterminal is predicted, making it po%ible, for the first time, for one to model the putative Fisinvettase complex in the continuing kfforts to unveil the molecular mechanism of invertase-, mediatedDNA recombination.