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BiochemistryTest（partone）NameStudentIDScoreIIIIIIIVTotalIWritetheStructuresandstatetheirapplicationsrespectivelyinBiochemistry.(5x4=20)NumberStructureApplications1dicyclohexylcarbodiimide多肽合成中氨基酸单体的活化试剂或缩水试剂。2cyanogenbromide特异性断裂甲硫氨酸C-端肽键3aspartylproteaseactivesite两个天冬氨酸构成天冬氨酸蛋白酶的活性位点。4ATP能量分子，磷酸基团供体5cAMP蛋白激酶A的活化信使IIMatchingQuestions(30x1=30)Usetheitemslistedinthetabletoanswerquestions1-15.Notalloftheanswerswillbeused.ABCDE1zymogenactivationphosphorylationkinasesligasephosphatases2evolutionarytreeconvergentdivergerntthalassemiasickle-cell3centrifugationionexchangeaffinitygelfiltrationprecipitation4synonymsanonymescleftinnersurface5competitivenoncompetitiveuncompetitivefeedbackfeedforward6turnovernumbercovalentbondsaltbridgevanderWaalsasparagine7cysteinehistidinearginineaspartatevaline8-carboxyglutamatetyrosinelysineserinehydroxyproline9ATPcAMPcGMPhomotropicheterotropic10primarysecondarytertiarynegativepositive11restrictionendonucleasesorthologsparalogsadultfetal12superoxideBohreffectSDsequencegladomainfingerprint13RamachandranSangerChargaffmyoglobinmetmyoglobin14isozymesorisoenzymesRibozymeallostericcooperativenoncooperative15stopped-flowRNAigenedisruptionZn2+Mg2+16reversetrancriptaseDNApolymeraseRNApolymeraseFe+2Fe+317geneexpressiongenetranscriptionmRNAstRNAsrRNAs18site-directedmutagenesiselagationreleaseexpressioncloning1.Someinvertedrepeatsequencesorpalindromicsequencescanbespecificallycleavedby____11A_________.2.Theprocessbywhichverydifferentevolutionarypathwaysleadtoaproteinwithsimilarfunctionisreferredtoas__2B___evolution.3.Undernormalconditions,theironofhemeinmyoglobinandhemoglobinisinthe___16D____oxidationstate.4.Carbondioxidereactswiththeaminoterminalgroupsofhemoglobintoformcarbamategroups,whichcarrya___10D___charge.5.Innormaladulthemoglobin,HbA,theβ6positionisaglutamateresidue,whereasinsickle-cellhemoglobin,HbS,itisa__7E___residue.6.TheT-stateofhemoglobinisstabilizedbyasaltbridgebetweenβ1Asp94andtheC-terminal____7B____oftheβ1chain.7.Intrypsin,thespecificitypocketcontainsa/an____7D____residuethatbindstoaminoacidresiduecarryingpositivechargeofthesubstrate.8.Inproteasessuchaspapain,a_____7A___residueisactivatedbyhydrogen-bondingtoahistidineresidue.9.Multipleformsofhomologousenzymeswithinthesameorganismthatcatalyzethesamereactionbutwithdifferentkineticpropertiesare____14A____.10.Theeffectsofsubstratesonallostericenzymesarereferredtoas_____9D_____effects.11.___9A___isthemostcommondonormoleculeinphosphorylationreactionscatalyzedbyproteinkinases.12.αhelices,βsheets,andturnsarereferredas__10B___structureofproteins.13.Thepurificationbasedontheattractionoftheproteinforaparticularchemicalgroupis___3C__chromatography.14.Theinformation-carryingintermediatesfortranslationare_17C_.15.ThepersonwhonotedtheA:TandG:CratiosinDNAareapproximately1:1is_13C_.16.Codonsthatspecifythesameaminoacidsaretermed__4A__.17.ApatternofDNAfragmentsbyrestrictiondigestionscanserveasa_12EofaparticularDNAmolecule.18.Glutaminesubstitutionwith__6E__isanexampleofaconservativeaminoacidchange.19.Thedecreasedproductionofoneofthesubunitsofhemoglobincancause_2D__.20.Twoαchainsandtwoγchainsareusedtoconstitute___11E___hemoglobin.21.Thekcatisoftenreferredtoasthe_6A__.22.EnzymesthatdonotobeyMichaelis-Mentenkineticsare_14C__.23.Theactivityofcarbonicanhydraserequiresmetalion__15D__.24.Wecanuse__18A___todeterminestructure/functionrelationshipsinenzymes.25.Weusuallyrefertheinhibitionofthefirststepbythefinalproductofthepathwayas__5D__inhibition.26.Activationofprothrombinrequiresmodifiedaminoacid__8A__ingladomain.27.TheenzymeusedtocreateDNAfromRNAis16A.28.Removalofphosphatesfromproteinsiscatalyzedbyprotein__1E__.29.Ω,βturnsandloopsareoftenfoundon4Eofproteins.30.Bloodclottingcascadesarecontrolledby__1A_.IIIFillintheBlanks(2x10=20)1.ThearomaticaminoacidsinproteinsarePhe,Tyr,andTrp.2.Duringproteinpurification,both测定比活性和回收率arenecessaryeachstep.3.Monoclonalantibodiesaremadeby生产抗体的B细胞与小鼠骨髓瘤细胞融合.4.Semi-conservativereplicationofDNAwasconfirmedbyM.MeselsonandF.W.Stahlexperiment(s).5.Distalhistidineinhemoglobindonateshydrogenbondtooxyhemoglobincomplex,whichinhibitstheoxidationoftheirontotheferricstate.6.α-hemoglobinisstabilizedbyAHSP.7.ThemeaningofVmaxcatalyzedbyanenzymeis所有活性位点都与底物结合时的反应初速度.8.Anaffinitylabelofanenzymeis有反应活性基团的底物类似物.9.ThecentralaminoacidresidueofthecatalytictriadinchymotrypsinisHis.10.Formedicaldiagnosis,LDH同工酶canbeusedasheartattackindicators.IVShort-AnswerQuestions（5x6=30）1.Oxygen-bindingcurvesbyhemoglobinareshowninFig1,whichindicatesthatO2affinityoftheproteinisregulatedbyO2,pH,CO2,and2,3-bisphosphateglycerol.Proposethemolecularmechanismstointerprettheoxygen-bindingcurveswiththeknowledgeyouhavelearntinclass.2.BrieflydescribeimmunoassayinBiochemistry.3.A10kbDNAfragmentdigestedwithdifferentrestrictionendonuclease(s)showninFig2.Pleasedrawapossiblerestrictionmapfortheoriginalfragment.4.Describestructuralalternationwhenthepeptidebondbetweenaminoacidresidues15and16iscleaved.5.HowtocreatehighdensityarraysofoligonucleotidesinAffymatrixCo?Whatcanbedoneif