近生理条件下DNA加速氧化修饰的铜锌超氧化物岐化酶聚集

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1、华中科技大学硕士学位论文近生理条件下DNA加速氧化修饰的铜-锌超氧化物岐化酶聚集姓名：殷俊申请学位级别：硕士专业：无机化学指导教师：刘长林20080605华中科技大学硕士学位论文AbstractAmyotrophiclateralsclerosis(ALS)isoneofthemostcommonadultneurodegenerativediseaseswithunknowncauses.Thestructuralandthemetal-bindingvariationsinCuZnsuperoxidedi

2、smutase(SOD1)mutantshavebeenlinkedtomanykindsofneurodegenerativediseases,inparticular,amyotrophiclateralsclerosis,duetotheiroxidativedamageandaggregation.Itiswellknownthatoxidation-inducedmodificationofSOD1canbeusedtomimicSOD1mutantsinvitrounderphysiologica

3、lconditions.TheaggregationbehaviorofSOD1wereexaminedunderoxidativeconditions,whichcouldmimictheeffectofmutationsandreflectthepracticalprocessdoneunderphysiologicalconditionstoahighextent.Thepresentstudyexaminedtheoxidation-inducedmodificationofSOD1inthepres

4、enceofascorbateandDNAcanbeusedtoacceleratetheformationofSOD1aggregatesbyaseriesofbiochemicalexperiments.Theoxidation-inducedmodificationofSOD1weretestedbySDS-PAGEand2-Delectrophoresis.TheresultsrevealthatthepIandthehydrophobicityofoxidativeSOD1areincreased.

5、TwoformsofDNAweretestedtotriggertheoxidativeSOD1aggregationbylightscatteringanddirectobservationsundervisiblelight,theoneformofDNAisdouble-strandedDNA(dsDNA),theotherformofDNAissingle-strandedDNA(ssDNA).TheresultsrevealthatdsDNAactsasatemplateforacceleratin

6、gtheformationofSOD1aggregatesandisincorporatedintoSOD1aggregates.AsignificantalterationinthepIandthehydrophobicityofSOD1causedbyoxidation,andtheenrichmentinSOD1alongdsDNAdoublestrandsaretwomainreasonsresponsibleforDNA-acceleratedSOD1aggregation.Theaccelerat

7、ioneffectofSOD1aggregationinvitrouponadditionofssDNAof24nucleotideswasexaminedunderphysiologicalconditions.ssDNAwastestedtotriggertheSOD1aggregationbylightscatteringanddirectobservationsundervisiblelight.TheresultsrevealthatssDNAcanacceleratetheformationofs

8、sDNA-SOD1aggregatemonomer,oligomericaggregate,microaggregate,andmacroaggregate.AlltheseresultsindicatethatthessDNA-acceleratedformationofinsolubleSOD1aggregatescanactasapotentialpathwaytoavoidaccumulat