生物化學Biochemistry生物化學1緒論大綱：課程主要內容及要求引子：生物化學的歷史和發展趨勢緒論2教材：《生物化學》，第三版，王鏡巖等主編，2002年，高教出版社參考書：1.Biochemistry,

JeremyM.Berg,JohnL.Tymoczko,LubertStryer5theditionW.H.FreemanandCompany2.Biochemistry,DonaldVoet,JudithG.Voet.3rdedition,2003,JohnWiley&Sons,Inc.NewYork.

教材：3生物化學（上）60學時其中上課時間：54學時生物化學（上）60學時4第一章緒論(3學時)（周叢照）第二章蛋白質(15學時)（周叢照）§2.120種氨基酸的結構和性質（2學時）§2.2蛋白質中的二級結構（1學時）§2.3蛋白質的高級結構（3學時）§2.4血紅蛋白和免疫球蛋白的結構與功能（3學時）§2.5維持蛋白質高級結構的作用力（1學時）§2.6蛋白質折疊和結構進化（2學時）§2.7蛋白質分離純化（3學時）

第三章核酸（6學時)§3.1核酸和核苷酸的結構、性質和功能（3學時）§3.2核酸的研究方法（3學時）第一章緒論(3學時)5第四章糖（3學時）§4.1糖的生物學作用（1學時）§4.2單糖和多糖（1學時）§4.3糖蛋白（1學時）第五章脂類和生物膜（6學時）§5.1脂的分類和性質（3學時）§5.2生物膜（2學時）§5.3膜蛋白（1學時）第四章糖（3學時）6第六章酶（15學時）§6.1酶的發展歷史、作用特征及催化機理(3學時)§6.2酶的分類及酶功能的多樣性（2學時）§6.3核酶（1學時）§6.4酶促反應動力學（3學時）§6.5酶的抑制作用（3學時）§6.6別構酶及其作用原理（3學時）

第七章維生素與激素（6學時）（周叢照）§7.1維生素與輔酶（3學時）§7.2激素及其作用原理（3學時）第六章酶（15學時）7生物化學（下）40學時其中上課時間36學時生物化學（下）40學時8第八章代謝總論（3學時）§8.1代謝及代謝途徑的相關概念（1學時）§8.1生物能學（2學時）第九章生物膜和物質運輸（3學時）§9.1物質跨膜運輸的方式（0.5學時）§9.2小分子物質的運輸（1學時）§9.3離子載體（1學時）§9.4生物大分子的跨膜運輸（0.5學時）第十章糖酵解（6學時）§10.1糖酵解概述（1學時）§10.2糖酵解的第一階段（2學時）§10.3糖酵解的第二階段（2學時）§10.4糖酵解的調控（1學時）第八章代謝總論（3學時）9第十一章檸檬酸循環（3學時）§11.1檸檬酸循環的準備階段（0.5學時）§11.2檸檬酸循環的反應機制（2學時）§11.3檸檬酸循環的調控（0.5學時）第十二章生物氧化（6學時）§12.1生物氧化基本概念和呼吸鏈（1.5學時）§12.2氧化還原電位和自由能變化（0.5學時）§12.2氧化磷酸化（4學時）第十三章光合作用（3學時）§13.1光合作用概況（1學時）§13.2光合磷酸化和CO2固定（2學時）第十一章檸檬酸循環（3學時）10第十四章糖原的分解和生物合成（3學時）§14.1糖原的降解和糖原的生物合成（2學時）§14.2糖原代謝的調控（1學時）第十五章脂肪酸代謝（3學時）第十六章氨基酸的分解代謝（3學時）第十七章核酸代謝（3學時）第十四章糖原的分解和生物合成（3學時）11WhatisBiochemistry?

thestudyofthosemoleculesusedandmanufacturedbylivingthings.WhatisBiochemistry?12Threeaspectsofbiochemistry:

1)Biochemistryisconcernedwithstructuralchemistry.Itseekstodeterminethestructuresofmoleculesfoundinlivingsystemsinordertounderstandstructure-functionrelationships.StructuralBiologyThreeaspectsofbiochemistry:132)Biochemistryisconcernedwithchemicalchange,thisisreflectedinthestudyofmetabolicpathwaysMetabolism2)Biochemistryisconcernedw143)BiochemistryisconcernedwithinformationwhichhasaccumulatedthroughevolutionandispreservedinDNA(orsometimesRNA).MolecularBiologyTheCentralDogma3)Biochemistryisconcernedw15BiochemistrythroughevolutionHowtobuildalifewithmolecules?OrTheMolecularDesignofLife

Biochemistrythroughevolution16胰島素由A、B兩個肽鏈組成。人胰島素(InsulinHuman)A鏈有21個氨基酸，B鏈有30個氨基酸，共51個氨基酸組成。其中A7(Cys)-B7(Cys)、A20(Cys)-B19(Cys)四個半胱氨酸中的巰基形成兩個二硫鍵，使A、B兩鏈連接起來。此外A鏈中A6(Cys)與A11(Cys)之間也存在一個二硫鍵。人工合成胰島素胰島素由A、B兩個肽鏈組成。人胰島素(InsulinHum171958年，中國科學院在王應睞、曹天欽、鄒承魯、鈕經義、沈昭文等先生的帶領下，提出了“世界上第一次用人工方法合成的蛋白質在中華人民共和國實現”的宏偉目標。1959年初，人工合成胰島素的工作全面展開。首先是由鈕經義、沈昭文、龔岳亭幾位研究人員組織有關人員解決了氨基酸的大量供應問題。

1965年，完成了結晶牛胰島素的合成，它有著極為深遠的意義。由于蛋白質和核酸兩類生物高分子有生命現象中所起的主要作用，人工合成了第一個具有生物活力的蛋白質，便突破了一般有機化合物領域到信息量集中的生物高分子領域之間的界限，在人類認識生命現象的漫長過程中邁出了重要的一步。合成胰島素工作的簡報發表于1965年《中國科學》。1958年，中國科學院在王應睞、曹天欽、鄒承魯、鈕經義、沈昭18Fourtransitionsthroughevolution:1,chemicals,micromolecules2,macrobiomolecules3,energy4,stressresponseFourtransitionsthroughevolu19Theevolutionofliferequiredaseriesoftransitions,beginningwiththegenerationoforganicmoleculesthatcouldserveasthebuildingblocksforcomplexbiomolecules.Theevolutionofliferequired20Thenextmajortransitionintheevolutionoflifewastheformationofreplicatingmolecules.EvolutionRequires:Reproduction,Variation,SelectivePressureThenextmajortransitionint21Variationwasintroducedbyanumberofmeans,fromsimplebasesubstitutionstotheduplicationofentiregenes.RNAappearstohavebeenanearlyreplicatingmolecule.Furthermore,someRNAmoleculespossesscatalyticactivity.However,therangeofreactionsthatRNAiscapableofcatalyzingislimited.Withtime,thecatalyticactivitywastransferredtoproteins,linearpolymersofthechemicallyversatileaminoacids.RNAdirectedthesynthesisoftheseproteinsandstilldoesinmodernorganismsthroughthedevelopmentofageneticcode,whichrelatesbasesequencetoaminoacidsequence.Eventually,RNAlostitsroleasthegenetothechemicallysimilarbutmorestablenucleicacidDNA.Inmodernorganisms,RNAstillservesasthelinkbetweenDNAandprotein.Variationwasintroducedbya22EnergyTransformationsAreNecessarytoSustainLivingSystemsATPservesasthecellularenergycurrencythatlinksenergy-yieldingreactionswithenergy-requiringreactions.ATPitselfisaproductoftheoxidationoffuelmolecules,suchasaminoacidsandsugars.Withtheevolutionofmembraneshydrophobicbarriersthatdelineatethebordersofcellsiongradientswererequiredtopreventosmoticcrises.ThesegradientswereformedattheexpenseofATPhydrolysis.Later,iongradientsgeneratedbylightortheoxidationoffuelmoleculeswereusedtosynthesizeATP.

EnergyTransformationsAreNec23CellsCanRespondtoChangesinTheirEnvironments

Thefinaltransitionwastheevolutionofsensingandsignalingmechanismsthatenabledacelltorespondtochangesinitsenvironment.Thesesignalingmechanismseventuallyledtocell-cellcommunication,whichallowedthedevelopmentofmore-complexorganisms.Therecordofmuchofwhathasoccurredsincetheformationofprimitiveorganismsiswritteninthegenomesofextantorganisms.(molecularevolution)CellsCanRespondtoChangesi24HistoryofBiochemistry(upto1982)HistoryofBiochemistry25

1835JonsBerzelius

chemicalcatalysis,usesamylase(淀粉酶)asanexample.1859CharlesDarwinpublishes

OntheOriginofSpecies.LouisPasteur

fermationcatalyzedbyenzymes,"essence"ofyeast.1865GregorMendelpublisheshis

theoryofgenetics.1869FredrickMeischerdiscovers

DNAincellnuclei.EduardandHansBuchnerextractsmaterielfromyeast,conversionofglucosetoalcohol.1835JonsBerzelius26

1914FritzLipmann,theroleofATPinenergymetabolism.1926JamesSumner,

crystallinejackbean(刀豆)urease,isaprotein.1926ThomasHuntMorganwrites

TheTheoryoftheGene.1934ArnoldBeckmandevelopesthefirstpHmeter.1937HansKrebsdiscoversthecitricacidcycle(TCAcycle).1941GeorgeBeadle&EdwardTatum,theone-gene,one-enzymehypothesis.OswaldAvery,ColinMacLeod,andMaclynMcCarthy

DNAisthegeneticmaterial.1914FritzLipmann,27

1950EdwinChargaff

A=T,G=C(Chargaff'srules).1952LinusPaulingandRobertCorey

α-helixandtheβ-pleatedsheet1953JamesWatsonandFrancesCrickthedoublehelixmodelofDNA.1953FredrickSangerthefirstaminoacidsequenceofaprotein(insulin).1956EarlSutherlandisolatescyclicAMP.1957MatthewMeselsonandFranklinStahl

semiconservativeDNAreplication.1950EdwinChargaff28

1960JohnKendrewandMaxPertuzobtain

thefirst3-Dstructureofproteins

(hemoglobinandmyoglobin).1960JeraldHuritzandSamuelWeissdiscover

RNApolymerase.1961FrancoisJacobandJaquesMonodpropound

theoperonmodelofgenecontrol.1963Jean-PierrreChanguex,F.Jacob,andJ.Monod

Allostericmodelforinhibitionofenzymes1964Severalgroups

Acrylamidegelelectrophoresisofproteinsisdeveloped1960JohnKendrewandMaxPe29MarshalNirenberg,H.GobindKhorana,andSeveroOchoacompletetheelucidationofthegeneticcode1965DavidPhillips

3-Dmodeloffirstenzyme(lysozyme)1965RobertHolleydetermines

thestructureofatransfer-aaRNA.1965JeromeVinograddiscovers

superhelicaltwisting.1968MarkPtashneandWalterGilbertidentify

thefirstrepressorgenes1969Firstsynthesisofanenzyme(ribonuclease).MarshalNirenberg,H.Gobind30

1970HamiltonSmithdiscovers

restrictionendonucleases.1970HowardTeminandDavidBaltimorediscover

reversetranscriptase.1973StanleyCohenandHerbertBoyerprepare

recombinantDNA.1974Sung-HouKim,duce

thefirstX-raystructureoftransferRNA.1977CesarMilsteindiscovershowtoproduce

monoclonalantibodies.1977AllanMaxamandWalterGilbertdevelop

achemistryforsequencingDNA.1977FredrickSanger,S.NicklenandA.R.Coulsondevelop

achemistryforsequencingDNA.1977PhillipSharpandRichardRobertsdiscover

introns(interveningsequences).1982AmzelLM,McKinneyM,NarayananP,PedersenPL

Firstx-raystructureofamembraneprotein(9?).1970HamiltonSmithdiscove311980sandearly1990sWonderfultimeforMolecularBiology1980sandearly1990s32HumanGenomeProject(1988-)CentralDogma(CrickF.1958)MolecularBiologyOmics:

MetabolomicsTranscriptomicsProteomics

StructuralGenomics

(1998-)

GenomicsHumanGenomeProject(1988-)Ce33TheeraofOmics:(1998-2003orlater?)Omics=Oh!Mix!(millenniumchaos?)TheeraofOmics:34Whatis

StructuralGenomics?Itisanapproachaimingatsolving

3-Dstructures

oftheproteinsencodedbyan

entiregenome.

KimSH.NatureStructuralBiology1998WhatisStructuralGenomics?Ki35OriginalgoalsofSG:1,toestablishacatalogue/libraryofallfoldscoveringtheentireproteinuniverse2,tohelptheannotationofsequencedgenomesOriginalgoalsofSG:36InternationalStructuralGenomicsProjectsUSAandCanada:(1998-)9NIHStructuralGenomicsCentersEU:France(3centers:Paris-Sud,Marseille,Strasbourg)Germany(StructuralGenomicsFactory)UK(2centers)(SPINE:StructuralProteomicsinEurope,2001-)Asia:Japan(RIKEN…)China(Yun-YuSHI;ZiheRAO…)(2002-)Korea:Israel:WeizmannStructuralProteomicsCenterInternationalStructuralGenom37Flowchart:1,Targetsselection:Bioinformatics2,PCRandcloning:MolecularBiology3,Proteinproduction:Biochemistry4,Datacollection(X-raydiffraction/NMR)5,Structuredetermination:StructuralBiology6,Functioninterpretation:GeneralBiologyFlowchart:38Corecharacteristics:Highthroughput(hundredstothousandssamples)Large-scale(milligramlevelofproteinsample)Multi-disciplineintegration(frommolecularbiologytostructuralbiology)Corecharacteristics:39highthroughput

parallelization(平行化)

miniaturization(微量化)

automation(自動化)highthroughput40cloningrobot:500-1000clones/3dayscrystallizationrobot:96x3drops/5minproteinsampleforcrystallizationtrialsinnanolitervolumes(<10nl)HansenCL.etal.PNAS;2002cloningrobot:500-1000clone41TagSize(aa)ApplicationSequenceorGenBankaccessionno.His6--10affinityHHHHHH(HH)T711affinityASMTGGQQMGRS15affinityKETAAAKFERQHMDSArg1,5,6affinityRRRRR(R)CBP26affinityKRRWKKNFIAVSAANRFKKISSSGALFLAG4or8affinityDYKDorDYKDDDDKStrep8affinityWSHPQFEKNusA491SET,affinityAccessionno.AAN82367MBP396SET,affinityAccessionno.AAC43128GST220SET,affinityAccessionno.AAB59203ZZ116SET,affinityAccessionno.M74186TrxA109SETAccessionno.AAC40210Gb156SETAccessionno.1MPEADsbADsbC208236SETSETAccessionno.P24991Accessionno.P21892Differenttagswillmakethedifference

TagSize(aa)ApplicationSequenc42DifferentE.colistrains(DE3series)

BL21(DE3)BL21-pLysSBL21-codon-plusTunerRosettaRosetta-pLysSGoldStarDifferentE.colistrains(DE343Co-expressionwithchaperones(DnaK-DnaJ-GrpEand/orGroEL-GroES)

Mainlyworksforthosepartiallysolubletargets

Co-purificationofchaperoneswiththetargetprotein

Re-aggregation/precipitationafterremovalofchaperonesCo-expressionwithchaperones44Eukaryoticexpressionsystems(post-translationalmodifications)PichiapastorisSaccharomycescerevisiaeInsect

cells(Sf9、SF21)Mammaliancells(CHOetal.)Virus-mediatedQuantityandCost-effectivityEukaryoticexpressionsystems45DNAshufflingSaturationmutagenesisError-pronePCR…Toimprove:

thesolubility,activityand/orstabilityReetz,PNAS;2004invitro/directedevolutionDNAshufflingReetz,PNAS;200446Proteinssamplesforcrystallizationtrials:1,pureandhomogenous2,stableinsolutionoflowsaltconcentration,properpHandreductant(BME,DTTorTCEP)3,athighconcentration(normally~10mg/ml)Proteinssamplesforcrystalli47HangingdropSittingdropHangingdropSittingdrop48《生物化學》簡介課件49ProteinsamplesforNMRdatacollection:1,doublelabelingwith13Cand15N2,nooligomerizationoraggregation3,stableforatleastoneweekatRTor4oC4,highexpressionlevel(cost-effective)ProteinsamplesforNMRdatac50Optimizationofxtals:1,abouthalfofthextalswillnotdiffractat3?orhigherresolution2,parameters:saltconcentration,buffersystem,pH,divalentortrivalentirons3,precipitants:PEGs,MPD,salt,alcohol,…4,ligandsorsubstrates/products/analogs5,proteinornucleicacidpartnersOptimizationofxtals:51Datacollection:X-raydiffraction:in-houseX-raygeneratorsynchrotronradiationacceleratorNMRspectrometry:500mHz,600mHz,800mHz900mHzDatacollection:52in-houseX-raygeneratorDatacollection:synchrotronradiationacceleratorin-houseX-raygenerator53

Assignmentoffunctionbasedonstructuralsimilarity

Reciprocalstimulationandvalidation:biochemicalassaysversusstructureanalyses

Structure-directeddrugdesign

Functioninterpretationandapplication:Assignmentoffunctionbased54

Assignmentoffunctionbasedonstructuralsimilarity3-DstructureDALIcomparisonCloseststructuresSimilarmolecularfunction?http://www.ebi.ac.uk/daliAssignmentoffunctionbased55HolmL,KaariainenS,RosenstromP,SchenkelA.(2008)SearchingproteinstructuredatabaseswithDaliLitev.3.

Bioinformatics24,2780-2781.

HolmL,KaariainenS,Rosenstr56

Reciprocalstimulationandvalidation:biochemicalassaysvs.structureanalyses

AnexampleoflicTfromBacillussubstilisReciprocalstimulationandva57LicTmutant(active)H207D/H269DLicTwt(inactive)ComparisonoflicT-wtandlicTmutantGraille*andZhou*etal.JBC2005vanTilbeurghetal.EMBOJ.2001LicTmutant(active)LicTwt(i58Yangetal.EMBOJ.20021122mRNA1122mRNAPKD=10μMKD=1μMCATPRD2PRD1RATCATRNAYangetal.EMBOJ.20021122mR59

Structure-directeddrugdesign

AnexampleofThy1fromThermotogamaritima

Thy1:thymidylatesynthase-complementingprotein

presentinarchaea,prokaryotes,viruses

NOTineukaryotesLesley,SAetal.PNAS;2002Structure-directeddrugdesig60Thy1-FAD-dUMPThy1-dUMP-HEPES

Thy1-FAD-dUMPThy1-dUMP-HEPES61谷胱甘肽硫轉移酶（GST）Biochem.J.(2001)360,1±16Xenobiotics（異型生物質，芳香族化合物）GSH+XenobioticsGS-Xenobiotics毒性較強溶解度低體內富集毒性較弱溶解度高解毒排毒谷胱甘肽硫轉移酶（GST）Biochem.J.(200162GST參與胞內氧化還原平衡維持，能防止芳香族有毒物質的體內富集，具有解毒功能。真核模式生物釀酒酵母中，有5個GST蛋白（Gtt1、Gtt2、Gto1、Gto2、Gto3），但是沒有一個的三維結構被解析，并且與已報道的序列相比，相似度較低。真菌界中的GST蛋白家族分類很混亂，并且目前沒有真菌的GST蛋白的三維結構被解析。我們希望通過對Gtt2的研究，闡述清楚生化功能和反應機制，并對真菌中GST的分類提供新的信息。Gtt2的研究意義GST參與胞內氧化還原平衡維持，能防止芳香族有毒物質的體內富63Gtt2的晶體Apo-form2.2埃GTS-boundform磺化谷胱甘肽，2.2埃GSH-boundform谷胱甘肽，2.1埃Gtt2的晶體Apo-formGTS-boundformG64Gtt2的結構溶液中為穩定二體，依靠靜電和疏水作用力維持每個單體含有兩個結構域，N端Trx-like和C端全螺旋結構域Gtt2的結構溶液中為穩定二體，依靠靜電和疏水作用力維持每個65底物結合位點主要作用方式：氫鍵主要來源：N端結構域的殘基底物結合位點主要作用方式：氫鍵66經典的催化殘基在Gtt2中缺失GSH的硫原子周圍既沒有酪氨酸，也沒有絲氨酸，更沒有半胱氨酸。經典的催化殘基在Gtt2中缺失GSH的硫原子周圍既沒有酪氨酸67經典的三種催化類型酪氨酸催化型絲氨酸催化型半胱氨酸催化型經典的三種催化類型酪氨酸催化型絲氨酸催化型半胱氨酸催化型68關鍵的催化殘基：晶體結構的證據磺化谷胱甘肽谷胱甘肽Apo-formSer129和His133穩定的水分子很重要關鍵的催化殘基：晶體結構的證據磺化谷胱甘肽谷胱甘肽Apo-f69關鍵的催化殘基：生化實驗的證據突變了水分子游離活性下降關鍵的催化殘基：生化實驗的證據突變了水分子游離活性下降70還原經典的催化殘基空間結構比對突變T24，G27偶然突變導致新的催化模式升高不變或下降還原經典的催化殘基空間結構比對突變T24，G27偶然突變導致71從結構上分析分析G27的微環境推測：空間位阻生化實驗驗證從結構上分析分析G27的微環境推測：空間位阻生化實驗驗證72生化試驗驗證GlyAlaSerCysPhe活性下降側鏈體積增大生化試驗驗證GlyAlaSerPhe活性下降側鏈體積增大73GST家族的生物信息學分析GST家族的生物信息學分析74首次定義了一支新的GST亞家族。該亞家族具有與經典模式（Tyr-，Ser-，Cys-type）不同的一種新的催化位點。Gtt2是首次報道的來源于真菌的GST蛋白結構，為目前混亂的真菌GST蛋白分類提供了新的思路。MaXXetal.EMBOReports2009首次定義了一支新的GST亞家族。75/pdb/home/home.do/pdb/home/h76Lastupdate:TuesdaySep01,2009

Lastupdate:TuesdaySep01,277X-rayLastupdate:TuesdaySep01,2009

X-rayLastupdate:TuesdaySep78NMRNMR79ElectronMicroscopyElectronMicroscopy80GrowthOfUniqueFoldsPerYearAsDefinedBySCOPGrowthOfUniqueFoldsPerYea81FutureorientationsofSG1,Reconstructionofmultiproteincomplexes(basedoninteractomics)2,Systematicallysolvingthe3-Dstructuresofmembraneproteins(achallengeofnoveltechniques)3,SystemsBiologyFutureorientationsofSG82Interactomes:1,Yeasttwo-hybrid2,TAP(tandemaffinitypurification)3,MassSpectrometry4,Co-IP(coimmunoprecipitation)5,PhagedisplayInteractomes:83OverexpresstheputativeproteincomplexinvivoorReconstructitinvitrofromtheindividualproteins

Solvethe3-DstructurebymeansofX-raycrystallographyCryo-ElectronMicroscopyElectroncrystallography(2DEM)ElectrontomographyOverexpresstheputativeprote84SystematicallyStructuretheMembraneProteins:

Abigchallenge!!PDB:59,790structures,updatedon2009/09/01/pdb/index.htmlSystematicallyStructuretheM85《生物化學》簡介課件86Uniqueproteins*indatabase=202.

Numberofco?rdinatefilesindatabase=551.

*Includesproteinsofsametypefromdifferentspecies.Forexample,photosyntheticreactioncentersfromR.viridisandR.sphaeroidesareconsideredunique.Lastdatabaseupdate:22Aug2009Uniqueproteins*indatabase=87《生物化學》簡介課件88SystemsBiology(系統生物學):1,Conceptually,anemergentfieldthataimsatsystem-levelunderstandingofbiologicalsystems.

()2,Inpractice,acombinationofomicapproaches,dataintegration,modelingandsyntheticbiology.

(Geatal.TIG,2003)SystemsBiology(系統生物學):89Hypothesis-drivenresearchinsystemsbiologyKitanoH.Science;2002Hypothesis-drivenresearchin90GenomicsStructuralGenomicsInteractomicsSystemsBiologyWordsSentencesLiteraturesDNAseqencesProteinFunctionsInteractionNetworksSystemsParagraphsIdeaoriginatedfromSalietalNature2003GenomicsStructuralGenomicsInt91MolecularBiologySystemsBiologyIntegration&ReconstructionReduction(molecularfunctions)MassiveData(GenBank,PDBetal.)DigitaltransparentsystemadreamtoberealizedMolecularBiologySystemsBiolo92生物化學Biochemistry生物化學93緒論大綱：課程主要內容及要求引子：生物化學的歷史和發展趨勢緒論94教材：《生物化學》，第三版，王鏡巖等主編，2002年，高教出版社參考書：1.Biochemistry,

JeremyM.Berg,JohnL.Tymoczko,LubertStryer5theditionW.H.FreemanandCompany2.Biochemistry,DonaldVoet,JudithG.Voet.3rdedition,2003,JohnWiley&Sons,Inc.NewYork.

教材：95生物化學（上）60學時其中上課時間：54學時生物化學（上）60學時96第一章緒論(3學時)（周叢照）第二章蛋白質(15學時)（周叢照）§2.120種氨基酸的結構和性質（2學時）§2.2蛋白質中的二級結構（1學時）§2.3蛋白質的高級結構（3學時）§2.4血紅蛋白和免疫球蛋白的結構與功能（3學時）§2.5維持蛋白質高級結構的作用力（1學時）§2.6蛋白質折疊和結構進化（2學時）§2.7蛋白質分離純化（3學時）

第三章核酸（6學時)§3.1核酸和核苷酸的結構、性質和功能（3學時）§3.2核酸的研究方法（3學時）第一章緒論(3學時)97第四章糖（3學時）§4.1糖的生物學作用（1學時）§4.2單糖和多糖（1學時）§4.3糖蛋白（1學時）第五章脂類和生物膜（6學時）§5.1脂的分類和性質（3學時）§5.2生物膜（2學時）§5.3膜蛋白（1學時）第四章糖（3學時）98第六章酶（15學時）§6.1酶的發展歷史、作用特征及催化機理(3學時)§6.2酶的分類及酶功能的多樣性（2學時）§6.3核酶（1學時）§6.4酶促反應動力學（3學時）§6.5酶的抑制作用（3學時）§6.6別構酶及其作用原理（3學時）

第七章維生素與激素（6學時）（周叢照）§7.1維生素與輔酶（3學時）§7.2激素及其作用原理（3學時）第六章酶（15學時）99生物化學（下）40學時其中上課時間36學時生物化學（下）40學時100第八章代謝總論（3學時）§8.1代謝及代謝途徑的相關概念（1學時）§8.1生物能學（2學時）第九章生物膜和物質運輸（3學時）§9.1物質跨膜運輸的方式（0.5學時）§9.2小分子物質的運輸（1學時）§9.3離子載體（1學時）§9.4生物大分子的跨膜運輸（0.5學時）第十章糖酵解（6學時）§10.1糖酵解概述（1學時）§10.2糖酵解的第一階段（2學時）§10.3糖酵解的第二階段（2學時）§10.4糖酵解的調控（1學時）第八章代謝總論（3學時）101第十一章檸檬酸循環（3學時）§11.1檸檬酸循環的準備階段（0.5學時）§11.2檸檬酸循環的反應機制（2學時）§11.3檸檬酸循環的調控（0.5學時）第十二章生物氧化（6學時）§12.1生物氧化基本概念和呼吸鏈（1.5學時）§12.2氧化還原電位和自由能變化（0.5學時）§12.2氧化磷酸化（4學時）第十三章光合作用（3學時）§13.1光合作用概況（1學時）§13.2光合磷酸化和CO2固定（2學時）第十一章檸檬酸循環（3學時）102第十四章糖原的分解和生物合成（3學時）§14.1糖原的降解和糖原的生物合成（2學時）§14.2糖原代謝的調控（1學時）第十五章脂肪酸代謝（3學時）第十六章氨基酸的分解代謝（3學時）第十七章核酸代謝（3學時）第十四章糖原的分解和生物合成（3學時）103WhatisBiochemistry?

thestudyofthosemoleculesusedandmanufacturedbylivingthings.WhatisBiochemistry?104Threeaspectsofbiochemistry:

1)Biochemistryisconcernedwithstructuralchemistry.Itseekstodeterminethestructuresofmoleculesfoundinlivingsystemsinordertounderstandstructure-functionrelationships.StructuralBiologyThreeaspectsofbiochemistry:1052)Biochemistryisconcernedwithchemicalchange,thisisreflectedinthestudyofmetabolicpathwaysMetabolism2)Biochemistryisconcernedw1063)BiochemistryisconcernedwithinformationwhichhasaccumulatedthroughevolutionandispreservedinDNA(orsometimesRNA).MolecularBiologyTheCentralDogma3)Biochemistryisconcernedw107BiochemistrythroughevolutionHowtobuildalifewithmolecules?OrTheMolecularDesignofLife

Biochemistrythroughevolution108胰島素由A、B兩個肽鏈組成。人胰島素(InsulinHuman)A鏈有21個氨基酸，B鏈有30個氨基酸，共51個氨基酸組成。其中A7(Cys)-B7(Cys)、A20(Cys)-B19(Cys)四個半胱氨酸中的巰基形成兩個二硫鍵，使A、B兩鏈連接起來。此外A鏈中A6(Cys)與A11(Cys)之間也存在一個二硫鍵。人工合成胰島素胰島素由A、B兩個肽鏈組成。人胰島素(InsulinHum1091958年，中國科學院在王應睞、曹天欽、鄒承魯、鈕經義、沈昭文等先生的帶領下，提出了“世界上第一次用人工方法合成的蛋白質在中華人民共和國實現”的宏偉目標。1959年初，人工合成胰島素的工作全面展開。首先是由鈕經義、沈昭文、龔岳亭幾位研究人員組織有關人員解決了氨基酸的大量供應問題。

1965年，完成了結晶牛胰島素的合成，它有著極為深遠的意義。由于蛋白質和核酸兩類生物高分子有生命現象中所起的主要作用，人工合成了第一個具有生物活力的蛋白質，便突破了一般有機化合物領域到信息量集中的生物高分子領域之間的界限，在人類認識生命現象的漫長過程中邁出了重要的一步。合成胰島素工作的簡報發表于1965年《中國科學》。1958年，中國科學院在王應睞、曹天欽、鄒承魯、鈕經義、沈昭110Fourtransitionsthroughevolution:1,chemicals,micromolecules2,macrobiomolecules3,energy4,stressresponseFourtransitionsthroughevolu111Theevolutionofliferequiredaseriesoftransitions,beginningwiththegenerationoforganicmoleculesthatcouldserveasthebuildingblocksforcomplexbiomolecules.Theevolutionofliferequired112Thenextmajortransitionintheevolutionoflifewastheformationofreplicatingmolecules.EvolutionRequires:Reproduction,Variation,SelectivePressureThenextmajortransitionint113Variationwasintroducedbyanumberofmeans,fromsimplebasesubstitutionstotheduplicationofentiregenes.RNAappearstohavebeenanearlyreplicatingmolecule.Furthermore,someRNAmoleculespossesscatalyticactivity.However,therangeofreactionsthatRNAiscapableofcatalyzingislimited.Withtime,thecatalyticactivitywastransferredtoproteins,linearpolymersofthechemicallyversatileaminoacids.RNAdirectedthesynthesisoftheseproteinsandstilldoesinmodernorganismsthroughthedevelopmentofageneticcode,whichrelatesbasesequencetoaminoacidsequence.Eventually,RNAlostitsroleasthegenetothechemicallysimilarbutmorestablenucleicacidDNA.Inmodernorganisms,RNAstillservesasthelinkbetweenDNAandprotein.Variationwasintroducedbya114EnergyTransformationsAreNecessarytoSustainLivingSystemsATPservesasthecellularenergycurrencythatlinksenergy-yieldingreactionswithenergy-requiringreactions.ATPitselfisaproductoftheoxidationoffuelmolecules,suchasaminoacidsandsugars.Withtheevolutionofmembraneshydrophobicbarriersthatdelineatethebordersofcellsiongradientswererequiredtopreventosmoticcrises.ThesegradientswereformedattheexpenseofATPhydrolysis.Later,iongradientsgeneratedbylightortheoxidationoffuelmoleculeswereusedtosynthesizeATP.

EnergyTransformationsAreNec115CellsCanRespondtoChangesinTheirEnvironments

Thefinaltransitionwastheevolutionofsensingandsignalingmechanismsthatenabledacelltorespondtochangesinitsenvironment.Thesesignalingmechanismseventuallyledtocell-cellcommunication,whichallowedthedevelopmentofmore-complexorganisms.Therecordofmuchofwhathasoccurredsincetheformationofprimitiveorganismsiswritteninthegenomesofextantorganisms.(molecularevolution)CellsCanRespondtoChangesi116HistoryofBiochemistry(upto1982)HistoryofBiochemistry117

1835JonsBerzelius

chemicalcatalysis,usesamylase(淀粉酶)asanexample.1859CharlesDarwinpublishes

OntheOriginofSpecies.LouisPasteur

fermationcatalyzedbyenzymes,"essence"ofyeast.1865GregorMendelpublisheshis

theoryofgenetics.1869FredrickMeischerdiscovers

DNAincellnuclei.EduardandHansBuchnerextractsmaterielfromyeast,conversionofglucosetoalcohol.1835JonsBerzelius118

1914FritzLipmann,theroleofATPinenergymetabolism.1926JamesSumner,

crystallinejackbean(刀豆)urease,isaprotein.1926ThomasHuntMorganwrites

TheTheoryoftheGene.1934ArnoldBeckmandevelopesthefirstpHmeter.1937HansKrebsdiscoversthecitricacidcycle(TCAcycle).1941GeorgeBeadle&EdwardTatum,theone-gene,one-enzymehypothesis.OswaldAvery,ColinMacLeod,andMaclynMcCarthy

DNAisthegeneticmaterial.1914FritzLipmann,119

1950EdwinChargaff

A=T,G=C(Chargaff'srules).1952LinusPaulingandRobertCorey

α-helixandtheβ-pleatedsheet1953JamesWatsonandFrancesCrickthedoublehelixmodelofDNA.1953FredrickSangerthefirstaminoacidsequenceofaprotein(insulin).1956EarlSutherlandisolatescyclicAMP.1957MatthewMeselsonandFranklinStahl

semiconservativeDNAreplication.