1

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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
-

(4) Lhlamln (d) carrles small nlLrogen-conLalnlng molecules
39
/=.I7;<=9
- hup://www.youLube.com/waLch?v=-cul?xc9Wko
- hup://www.youLube.com/waLch?v=lgxnP087!lk&feaLure=relaLed
40