3-, 456 1 -"22%27& 8"9:.&7;<= - rocess ln whlch cells consume C 2 and produce CC 2 - rovldes more energy (A1) from glucose Lhan glycolysls - Also capLures energy sLored ln llplds and amlno aclds - Lvoluuonary orlgln: developed abouL 2.3 bllllon years ago - used by anlmals, planLs, and many mlcroorganlsms - Cccurs ln Lhree ma[or sLages: - aceLyl CoA producuon (breakdown amlno aclds, glucose, fauy aclds) - aceLyl CoA oxldauon (clLrlc acld cycle) - elecLron Lransfer and oxldauve phosphorylauon 2 2 8"9:.&7;<=* >$7?" 4 /#"$12@-</ A&<0%#;<= CeneraLes some: A1, nAuP, lAuP 2 3 8"9:.&7;<=* >$7?" ( /#"$12@-</ <B.07;<= CeneraLes more nAuP, lAuP 2 ,
and one C1 4 3 8"9:.&7;<=* >$7?" C DB.07;E" A,<9:,<&127;<= CeneraLes a loL of A1 3 +," -.$&.# /#.0 -1#2" - under aeroblc condluons, Lhe pyruvaLe produced by glycolysls ls oxldlzed Lo P 2 C and CC 2 - 1he clLrlc acld cycle ls a hub ln meLabollsm, wlLh degradauve paLhways leadlng ln and anabollc paLhways leadlng ouL. 6 4 >$7?" F* -<=E"&9.<= <G A1&%E7$" $< /#"$12@-</ - neL 8eacuon: - Cxldauve decarboxylauon of pyruvaLe - llrsL carbons of glucose Lo be fully oxldlzed - CaLalyzed by Lhe pyruvaLe dehydrogenase complex - 8equlres 3 coenzymes - llpoyllyslne, and lAu are prosLheuc groups - 1, nAu + and CoA-SP are co-subsLraLes 7 A1&%E7$" H",10&<?"=79" -<I:2"B 3AH-6 - uC ls a large (up Lo 4) JH7) muluenzyme complex - pyruvaLe dehydrogenase (L 1 ) - Coenzyme= 1 - dlhydrollpoyl LransaceLylase (L 2 ) - Coenzymes=llpolc acld, CoASP - dlhydrollpoyl dehydrogenase (L 3 ) - Coenzymes= lAu, nAu + - Complex= 24 L 2 +24 L 1 dlmers+ 12 L 3 dlmers - AdvanLages of muluenzyme complexes: ! shorL dlsLance beLween caLalyuc slLes allows channellng of subsLraLes from one caLalyuc slLe Lo anoLher. ! channellng mlnlmlzes slde reacuons ! regulauon of acuvlLy of one subunlL aecLs Lhe enure complex 8 5 DE"&722 8"7#;<= <G AH- 9 >$&%#$%&" <G -<"=K1I" / - Coenzymes are noL a permanenL parL of Lhe enzymes' sLrucLure. - 1hey assoclaLe, fulll a funcuon, and dlssoclaLe - 1he funcuon of CoA ls Lo accepL and carry aceLyl groups 10 6 >$&%#$%&" <G !.:<12219.=" - rosLheuc groups are sLrongly bound Lo Lhe proLeln - 1he llpolc acld ls covalenLly llnked Lo Lhe enzyme vla a lyslne resldue 11 DE"&722 8"7#;<= <G AH- 12 7 >"L%"=#" <G ME"=$9 .= DB.07;E" H"#7&N<B127;<= <G A1&%E7$" Lnzyme 1 - >$": 4* uecarboxylauon of pyruvaLe Lo an aldehyde - >$": (* Cxldauon of aldehyde Lo a carboxyllc acld ! LlecLrons reduce llpoamlde and form a LhloesLer Lnzyme 2 - >$": C* lormauon of aceLyl-CoA (producL 1) Lnzyme 3 - >$": O* 8eoxldauon of Lhe llpoamlde cofacLor - >$": P* 8egenerauon of Lhe oxldlzed lAu cofacLor ! lormlng nAuP (producL 2) 13 -<=$&<2 <G $," AH- - 1hls complex ls sLrongly lnhlblLed by A1, aceLyl-CoA, and nAuP. 1hey are all lndlcaLors of adequaLe energy levels. - AllosLerlc lnhlbluon ls enhanced by Lhe presence of long- chaln fauy aclds (an alLernauve fuel source). - AM, CoA, and nAu + allosLerlcally acuvaLe Lhe complex. 1hese are all lndlcaLors of lnadequaLe energy levels. - 1he complex ls also regulaLed by proLeln modlcauon. 8everslble phosphorylauon of L 1 lnhlblLs Lhe complex. - 1he klnase responslble for Lhls reacuon ls allosLerlcally acuvaLed by A1. 14 8 +," -.$&.# /#.0 -1#2" 3-/-6 13 -.$&.# /#.0 -1#2" DE"&E."Q - ln each Lurn of Lhe cycle, one molecule of aceLaLe enLers by condenslng wlLh oxaloaceLaLe Lo form clLraLe, Lwo molecules of CC 2 leave, and oxaloaceLaLe ls regeneraLed. - Slnce oxaloaceLaLe ls regeneraLed, ln Lheory only one molecule of Lhls compound ls needed Lo keep Lhe cycle golng. - lour of Lhe elghL sLeps are oxldauons, produclng nAuP or lAuP 2 . - varlous lnLermedlaLes can be slphoned o for oLher meLabollc paLhways and Lhey can be replenlshed by a varleLy of anaplerouc reacuons. - 1he enure seL of reacuons Lakes place ln Lhe mlLochondrla. MlLochondrla also conLaln Lhe enzymes for oxldauve phosphorylauon and for oxldauon of fauy aclds and amlno aclds Lo aceLyl-CoA. 16 9 +," -.$&.# /#.0 -1#2" 3-/-6 17 >$": 4* R<&I7;<= <G -.$&7$" 18 10 >$": (* R<&I7;<= <G F9<#.$&7$" - AconlLase caLalyzes Lhe formauon of lsoclLraLe from clLraLe, Lhrough Lhe lnLermedlary formauon of cls-aconlLaLe. - AconlLase conLalns an lron-sulfur cenLer whlch acLs ln blndlng of subsLraLe and ln caLalyuc addluon of P 2 C. - Addluon of P 2 C Lo Lhe lnLermedlaLe cls-aconlLaLe could produce elLher clLraLe or lsoclLraLe. - 1he reacuon ls pulled Loward formauon of lsoclLraLe, agalnsL free energy, because lsoclLraLe ls rapldly consumed ln Lhe nexL sLep of Lhe cycle. 19 20 11 >$": C* DB.07;<= <G F9<#.$&7$" - lsoclLraLe ls oxldlzed Lo !-keLogluLaraLe and CC 2 by Lhe enzyme lsoclLraLe dehydrogenase. - 1hls reacuon ls hlghly favorable and produces nAuP (or nAuP) plus P + . - 1he nAu-dependenL enzyme serves ln Lhe clLrlc acld cycle. 1he nAu + enzyme ls found ln Lhe cyLosol and Lhe mlLochondrla and ls probably requlred for Lhe generauon of nAuP ln Lhese comparLmenLs. 21 >$": O* DB.07;<= <G !@S"$<?2%$7&7$" - 1hls reacuon produces nAuP, ls hlghly favorable (-33.3 k!/mol), and conserves Lhe energy of oxldauon ln Lhe LhloesLer bond of succlnyl-CoA. - 1he reacuon of Lhls complex ls almosL ldenucal Lo LhaL of Lhe pyruvaLe dehydrogenase complex and Lhese Lwo proLelns are undoubLedly evoluuonarlly relaLed. - L 2 and L 3 are almosL ldenucal beLween Lhe Lwo complexes, whlle L 1 dlers and provldes subsLraLe speclclLy. 22 12 >$": P* >%##.=12@-</ >1=$,"$79" - Succlnyl-CoA synLheLase converLs succlnyl-CoA Lo succlnaLe, produclng C1 (or A1) from Cu (or Au) and ! ln Lhe process. - 1he enzyme ls LranslenLly phosphorylaLed on a hlsudlne resldue durlng Lhls reacuon. - Succlnyl-CoA synLheLase has Lwo subunlLs. 1he !-subunlL conLalns Lhe phosphorylaLed Pls resldue and blnds CoA. 1he "-subunlL confers speclclLy for Au or Cu. - now LhaL we have a four carbon compound agaln, we need Lo perform a few oxldauons ln order Lo generaLe oxaloaceLaLe. 23 24 13 >$": 5* >%##.=7$" H",10&<?"=79" - SucclnaLe ls oxldlzed Lo fumaraLe by Lhe avoproLeln succlnaLe dehydrogenase, produclng lAuP 2 . - SucclnaLe dehydrogenase ls a membrane bound proLeln, Lhe only one ln Lhe CAC cycle. - LlecLrons pass from succlnaLe Lo lAu and Lhen Lhrough Lhree lron-sulfur cenLers before enLerlng Lhe oxldauve phosphorylauon paLhway. - LlecLron ow from Lhls reacuon ylelds 1.3 A1 molecules per palr of elecLrons. - MalonaLe ls a sLrong compeuuve lnhlblLor of Lhls enzyme and blocks Lhe cycle. 23 >$": T* R%I7&79" - lumarase caLalyzes Lhe reverslble hydrauon of fumaraLe Lo malaLe, uslng a carbanlon Lransluon sLaLe. - 1hls enzyme ls hlghly sLereospeclc, hydraung Lhe Lrans double bond of fumaraLe buL noL Lhe cls double bond of maleaLe. - ln Lhe reverse dlrecuon only L-malaLe ls a subsLraLe, noL Lhe u-lsomer. 26 14 >$": U* DB.07;<= <G J727$" - L-malaLe dehydrogenase oxldlzes malaLe Lo oxaloaceLaLe, regeneraung Lhe sLarung compound and produclng nAuP. - 1hls reacuon ls unfavorable under sLandard condluons (29.7 k!/mol), buL oxaloaceLaLe ls kepL aL a low concenLrauon (<10 -6 M), pulllng Lhe reacuon forward. 27 M="&?1 -<=9"&E7;<= - Lnergy released durlng oxldauon ls conserved ln Lhe reducuon of 3 nAu + and one lAu, as well as Lhe producuon of one A1 (C1). - 1he Lwo carbons LhaL exlL as CC 2 are noL Lhe same carbons LhaL enLer as aceLaLe. - ln oxldauve phosphorylauon, Lhe elecLrons from nAuP produce 2.3 A1 and Lhe elecLrons from lAuP 2 produce 1.3 A1. - As many as 32 molecules of A1 are obLalned per glucose molecule. 28 15 D$,"& 8<2"9 <G $," +-/ -1#2" - Cxldauon of aceLaLe appears Lo be a compllcaLed process. - lL probably evolved under anaeroblc condluons and Lhen conunued Lo provlde a selecuve advanLage. Some modern anaeroblc organlsms have an lncompleLe clLrlc acld cycle, whlch lacks !-keLogluLaraLe dehydrogenase, whlch Lhey use as a source of blosynLheuc precursors. - 1he role of Lhe clLrlc acld cycle ls noL conned Lo oxldauon of aceLaLe, lL serves as Lhe hub of lnLermedlary meLabollsm. - lnLermedlaLes such as oxaloaceLaLe and !-keLogluLaraLe can be drawn ouL of Lhe cycle and used as precursors ln blosynLheuc paLhways of mosL amlno aclds. - Succlnyl-CoA serves as a precursor of Lhe porphyrln rlng of heme groups. - CxaloaceLaLe ls also a precursor ln glucose synLhesls. 29 30 16 /=7:2"&<;# 8"7#;<=9 - lnLermedlaLes ln Lhe clLrlc acld cycle are replenlshed by anaplerouc reacuons. - 1he concenLrauons of all of Lhe lnLermedlaLes ln Lhe cycle remaln almosL consLanL. - 1hese anaplerouc reacuons generally produce elLher oxaloaceLaLe or malaLe. - ln Lhe kldney and llver, pyruvaLe carboxylase forms oxaloaceLaLe from pyruvaLe and CC 2 . - yruvaLe carboxylase ls sLrongly sumulaLed by aceLyl-CoA. 31 32 17 8"?%27;<= <G $," -.$&.# /#.0 -1#2" 1. 1he clLrlc acld cycle ls under ughL regulauon aL Lwo levels: - 1he converslon of pyruvaLe Lo aceLyl-CoA (Lhe pyruvaLe dehydrogenase complex). - 1he enLry of aceLyl-CoA lnLo Lhe cycle (clLraLe synLhase). 2. 1he cycle ls also regulaLed aL Lhe lsoclLraLe dehydrogenase and !-keLogluLaraLe dehydrogenase reacuons. 33 8"?%27;<= <G A1&%E7$" H",10&<?"=79" -<I:2"B - lnhlblLed by hlgh energy lndlcaLors - AcuvaLed by low energy lndlcaLors CovalenL modlcauon: -phosphaLase removes l from L1 (acuve). AcuvaLed by calclum - klnase adds l Lo L1 (lnacuve). AcuvaLed by nAuP, aceLyl-CoA 34 18 -<=$&<2 7$ MB"&?<=.# >$":9 - lnhlbluon of clLraLe synLhase by A1 ls relleved by Au. - ln verLebraLe muscle, Ca 2+
acuvaLes lsoclLraLe dehydrogenase and !- keLogluLaraLe dehydrogenase. - 1he raLes of glycolysls and Lhe clLrlc acld cycle are lnLegraLed so LhaL boLh are operaung aL Lhe same raLe. 33 >%II7&.K" $," -/- aLhway: ClLrlc Acld Cycle urpose: aLhways leadlng ln: aLhways leadlng ouL: Locauon: Molecules needed for lnpuL: CuLpuL: AcuvaLors: lnhlblLors: 36 19 >%II7&.K" $," -/- aLhway: ClLrlc Acld Cycle urpose: aLhways leadlng ln: aLhways leadlng ouL: Locauon: Molecules needed for lnpuL: CuLpuL: AcuvaLors: lnhlblLors: 37 Generate energy in the form of NADH, FADH2, and GTP by oxidizing acetate Mainly glycolysis, fatty acid oxidation, amino acid degradation NADH, FADH 2 ! Oxidative phosphorylation (where ATP is made) Other intermediates ! anabolic pathways Mitochondrial matrix Acetyl-CoA, oxaloacetate (recycled) 2CO 2 , 3 NADH, 1FADH 2
AMP, Calcium, NAD, CoA ATP, Acetyl-CoA, NADH 8"7#;<=9 <G $," AHV -<I:2"B 7=0 -/- uP complex and CAC 8eacuons 1 ! 8 LlsL ouL for each: SubsLraLe(s) ! roducL(s) CofacLors (lf any) lmporLanL feaLures of Lhe mechanlsm lnhlblLors/acuvaLors 38 20 !"7&=.=? MB"&#.9" 3( :$96 MaLch Lhe cofacLor wlLh lLs funcuon ln Lhe clLrlc acld cycle. A glven funcuon may be used more Lhan once or noL aL all. CofacLor luncuon (1) nAu + /nAuP (a) carrles C 2
(2) lAu/lAuP 2 (b) carrles small carbon-conLalnlng molecules (3) CoA (c) carrles e -