江浙蝮蛇毒IX因子結合蛋白（AHP IX-bp）和尖吻蝮蛇毒鋅金屬蛋白酶AaH IV自降解產(chǎn)物D-C-motif(acucetin)的結構和功能關系研究.pdf

1、中國科學技術大學博士學位論文江浙蝮蛇毒IX因子結合蛋白（AHPIXbp）和尖吻蝮蛇毒鋅金屬蛋白酶AaHIV自降解產(chǎn)物DCmotif(acucetin)的結構和功能關系研究姓名：臧建業(yè)申請學位級別：博士專業(yè)：生物化學與分子生物學指導教師：牛立文滕脈坤20030701博1：論文摘要ABSTRACT1Purification，partialcharacterization，cDNAsequencing。crystallizationandst

2、ructuredeterminationofAHPIXbp，anewcoagulationfactorIXbindingproteinfromtheAgkistrodonhalysPallasvenomAnewcoagulationfactorIXbindingprotein，AHPIXbp，hasbeenpurifiedfromAgkistrodonhalysPallasvenomandestimatedtobeaABheterodi

3、merofabout25kDaconsistingoftwochains(A—chainof155kDaandBchainof15kDa)linkedbydisulfidebond(s)NterminalsequenceandpartialcDNAsequenceofAHPIX—bpwasdeterminedandalignedwithCtypelectin—likeproteinsTheproteinhasahighsequences

4、imilaritytosomeofthesnake—venomCtypelectinlikeproteinsAHPIX—bpcouldbindtocoagulationfactorIXbutnottocoagulationfactorXSoAHPIXbpisanewcoagulationfactorIXbindingproteinpurifiedfromsnakevenomAlignmentofthededucedaminoacidse

5、quenceofAHPIXbpwithotherSnakevenomcoagulationfactorIX／XbindingproteinsshowsthatthemajordifferencesoftheseproteinsfoCUSontheregionsofA56A67MoreoveEAHPIX；bDshowsthebindingtocoagulationfactorIXinbothzincion—dependentandpH—d

6、ependentmannersTheafnnitybetweenAHPIXbpandcoagulationfactorIXishigherunderneutralorweaklyalkalineconditionsthanthatunderweaklyacidicconditionsToiIluminatethestructuraIbasisofthiskindofbinding，singlecrystalsofAHPIXbpgrown

7、atpH65andpH75conditionswereobtainedanddiffractXraysto2OAand18AresolutionrespectivelyBothcrystalsareisomorphousandbelongtothespacegroupP1；onlyoneABheterodimerisinthewholeunitcellThestructuresofAHPIXbDatpH65andpH75havebeen

8、determinedto20Aand18AbythemethodofmolecularreplacementTheoveralIstructuresatofbothpHarealmostthesamewithrmsdeviationof0122AButHisB61iSobservedtotaketwoconformationsintheIowerpHstructureformThisconformationalchangemayindu

9、cethebiologicalactivitychange2PartiaICharacterizationandCrystallizationofAcucetin，AProteinContainingBothDisintegrinlikeandCysteineRichDomainsReleasedbyAutoproteolysisofAPⅢtypeMetalloproteinaseAaHIVfromAgkistrodonacutusVe