Photosynthesis

The Dark Reactions of

Photosynthesis, Assimilation of
Carbon Dioxide And The
Calvin Cycle
C
3
, C
4
and CAM. Re!lation of
The Activity of Photosynthesis
The "iht Reactions of
Photosynthesis
The Photosynthetic Membrane
"iterat!re
The observation that a #illo# that has been c!ltivated in a container for five years #ith
eno!h #aterin ained more than half a centner #eiht altho!h only t#o o!nces of the
container$s soil #ere lost oes back to %.&. van '("M)*T +,-.. / ,0441. The &ritish
nat!ral scientist 2. 'A"(2 +,0.. / ,.0,1 !nderstood that air and liht are necessary for
the n!trition of reen 3lants. &!t it #as not before the com3osition of air o!t of different
ases became kno#n that their sinificance for 3lant n!trition #as st!dies. 4n ,..,
observed %. PR4(2T"(5 +,.33 / ,6741, one of the discoverers of oxyen, that reen
3lants ive off oxyen and th!s im3rove the air.
The 3riest %. 2(*(&4(R +,.48 / ,6791 from :eneva discovered that the reeneration of
the air is based on the !se of $fixed air$ +carbon dioxide1. These observations #ere
confirmed and broadened by st!dies of the D!tch doctor %. 4*:(*');2< +,.37 / ,.991
#ho reconi=ed both the meanin of liht and the fact that the #hole carbon contained in
3lants is of atmos3heric oriin. 'e, too, conceived that 3lants take !3 small amo!nts of
oxyen at niht or in the shado# and ive off carbon dioxide. 4n ,674 discovered Th. des
2A;22;R( +,.0. / ,64-1 from :eneva that the 3lants$ increase in #eiht cannot solely
be ca!sed by the !3take of carbon and minerals, b!t is based on the bindin of the #ater
com3onents, too.
4n ,694 constr!cted T. >. (*:("MA** +,643 / ,9791 a adet o!t of a modified
microsco3e condenser that allo#ed him to ex3ose 3arts of 3hotosynthetically active cells
+of the reen ala Spirogyra1 to a thin ray of liht. 'is aim #as to discover #hich
com3onents of the cell f!nctioned as liht rece3tors. To meas!re the oxyen 3rod!ction,
he dis3ersed the thread/like Spirogyra in a bacteria/containin s!s3ension. >henever
3arts of the chloro3last #ere ill!minated, did the bacteria concentrate in this area +#here
oxyen #as available1. The ill!mination of other 3arts of the cell res!lted in no s!ch
areations.
4n an earlier st!dy did he s3lit #hite liht into its s3ectral com3onents !sin a 3rism. 'e
then ill!minated a reen ala, Chladophora, #ith this s3ectr!m. 4n contrast to Spirogyra
are the Chladophora cells com3letely and evenly filled by the chloro3last. 'e observed
that the bacteria acc!m!lated mainly in the bl!e and red liht. A first action s3ectr!m of
3hotosynthesis #as th!s yielded. 4t resembles ro!hly the absor3tion s3ectra of
chloro3hyll a and b.
%. v. 2AC'2 +,638 / ,69.1 co!ld finally 3rove that chloro3hyll is involved in
3hotosynthesis. 4n addition did he sho# that starch is 3rod!ced in chloro3lasts as a res!lt
of the 3hotosynthetic activities.
These res!lts are in accord #ith the first la# of thermodynamics, #hose discoverer %. R.
MA5(R 3ost!lated already in ,648 that 3lants take !3 enery in the form of liht and
that they transform it into another, a chemical state of enery. &ased on this ass!m3tion
#as the reaction e?!ation
0 carbon dioxide @ 0 #ater A (chlorophyll) A l!cose @ 0 oxyen
form!lated.
%.v. "4(&4: ass!med that the oxyen stems from the breakdo#n of the carbon dioxide.
This idea #as !ncritically acce3ted by the 3lant 3hysioloists of the late ,9
th
and the early
87
th
cent!ry +2AC'2, PB(BB(R, %)2T and others1 altho!h M. %. 2C'"(4D(* had as
soon as ,648 reali=ed that
,. l!cose is 3rod!ced as a res!lt of 3hotosynthesis +and he #as closer to
reality than 2AC'2 #as later1 and that
8. it is very likely that it is #ater that is broken do#n. 'e #roteC
D4t is #ell/kno#n that C)
8
is amon the most stabile com3o!nds and
that no chemical #ay of breakin it do#n is kno#n #hile '
8
) is very
easily broken do#n.... and it does therefore seem likely that the 84 '
8

of the 84 '
8
) are combined #ith the ,8 C)
8
.D
+fromC :r!nd=Ee der #issenschaftlichen &otanik1. 2C'"(4D(*$s e?!ations contain all
reaction com3o!nds in do!ble n!mbers. 'e ives C
,8
'
84
)
,8
as the form!la for l!cose.
4t #as soon reali=ed that the reaction e?!ation above is a sim3lification and that
3hotosynthesis consists of a n!mber of 3artial 3rocesses.
B. B. &"ACFMA* and :. ". C. MAT':(" +,97-, ;niversity of Cambride, :reat
&ritain1 #ere amon the first to st!dy this to3ic systematically. They c!ltivated 3lants
!nder different b!t controlled carbon dioxide concentrations, different liht intensities
and different tem3erat!res and they noted the effects of these 3arameters on the rate of
3hotosynthesis. T#o decisive as3ects #ere revealed. ;nder stron liht and limited
amo!nts of carbon dioxide is the rate of 3hotosynthesis de3endent on the tem3erat!re.
This sho#s that the carbon dioxide fixation is based on normal, tem3erat!re/de3endent
biochemical reactions. ;nder carbon dioxide excess and too little liht #as no
tem3erat!re/de3endence fo!nd. This hints at the fact that the liht/ind!ced reactions are
inde3endent of the tem3erat!re. This statement a33lies to all 3hotochemical reactions.
4n ,98- 3!t ). >AR&;R: +Faiser/>ilhelm/4nstit!t Glater Max/Planck/4nstit!tH fEr
<ell3hysioloie at &erlin/Dahlem1 the res!lts of &"ACFMA* do#n to the existence of
t#o classes of 3hotosynthetic reactionsC the liht and the dark reactions.
D!rin the thirties analy=ed C. &. van *4(" +2tanford ;niversity1 the 3hotosynthesis of a
n!mber of 3!r3le bacteria. 4n addition to carbon dioxide do these bacteria need hydroen
s!l3hide for 3hotosynthesis. van *4(" #as able to determine
0 C)
8
@ ,8 '
8
2 A (light) A C
0
'
l8
)
0
@ ,8 2 @ 0 '
8
)
as the reaction$s e?!ation. &ased on it did he extra3olate a eneral e?!ation of
3hotosynthesisC
C)
8
@ 8 '
8
I A (light) A +C'
8
)1 @ '
8
) @ 8 I
Accordin to this e?!ation is 3hotosynthesis a redox reaction #ith '
8
I as the electron
donator +the oxydi=able s!bstance1. 4n the case of reen 3lants is it '
8
) and this means
that not the carbon dioxide b!t the #ater is broken do#n.
A first ex3erimental 3rove that the oxyen develo3ed d!rin the 3hotosynthesis of reen
3lants stems indeed from #ater #as delivered by the &ritish 3hysioloist R. '4"". 'e
detected that isolated chloro3lasts ive off oxyen in the 3resence of !nnat!ral red!cin
aents like iron oxalate, ferricyanide or ben=o?!inone after ex3os!re to liht. The
reaction #ent do#n in literat!re as the '4""/reactionC
8 '
8
) @ 8 A A (light, chloroplasts) A 8 A'
8
@ )
8

#here A is the electron acce3tor. 4f A J Be
444
, then is
8 '
8
) @ 4 Be
444
A (light, chloroplasts) A 4 Be
44
@ )
8
@ 4 '
@

The 3rocess is linked to a 3hotolytic breakdo#n of #ater that 3recede the red!ctionn of
Be
444
.
4 '
8
) A (light, chloroplasts) A 4 '
@
@ 4 )'
/

This sho#s that
oxyen can also be set free in the absence of carbon dioxide,
the oxyen 3rod!ced stems from the breakdo#n of #ater,
isolated chloro3lasts are able to 3erform at least 3artial 3rocesses of 3hotosynthesis.
The statement that the oxyen 3rod!ced d!rin 3hotosynthesis stems only from the
breakdo#n of #ater #as confirmed by 2. M. R;&(*, M. RA*DA"", M. FAM(* and
%. ". '5D( in ,94, after the isoto3e techni?!e had fo!nd its #ay to biochemistry. They
co!ld sho#n that a s!s3ension of Chlorella ro#n in '
8
,6
), ives off ,6)
8
, after liht
ex3os!re. 2hortly after#ards confirmed 2. M. R;&(* and his collaborators the 3ost!late
of ). >AR&;R: that the fixation of carbon dioxide is enery cons!min b!t
inde3endent of liht. 4n addition co!ld (. RACF(R +Cornell ;niversity, 4thaca, *. 5.1
3rove that liht can be re3laced by the addition of enery/rich com3o!nds.
The Dark Reactions of Photosynthesis, Assimilation of Carbon
Dioxide And The CALVIN Cycle.
D!e to the !se of isoto3es #ere M. CA"K* and his collaborators at the ;niversity of
California, &erkeley able to reveal com3letely the reactions takin 3lace d!rin the
incor3oration of carbon dioxide into carbohydrates in the relatively short 3eriod from
,940 / ,9-3. The ?!ick s!ccess #as based on the !se of sensitive methods +t#o/
dimensional 3a3er chromatora3hy, a!toradiora3hy1, a s!itable s3ecimen and the ra3id
3roresses of en=yme biochemistry. C!lt!res of the sinle/celled reen ala Chlorella
pyrenoidosa +that #as introd!ced to 3hotosynthetic st!dies in ,9,9 by ). >AR&;R:1
#ere s!33lied #ith liht and an even stream of air containin
,8
C)
8
.
At a iven time +tJ 71 #as
,4
C)
8
added to the stream of air for a short time. 4t #as
ass!med that the labelled carbon dioxide molec!les #ere s!ccessively incor3orated into
intermediates of the carbohydrate synthesis. After 3, - etc. seconds #ere the ex3eriments
sto33ed by addin boilin alcohol and the ne#ly 3rod!ced
,4
C/labelled intermediates
#ere se3arated and identified by 3a3er chromatora3hy.
,. The first stable com3o!nd that #as labelled radioactively already after 3 seconds
#as 3/3hos3holycerate +3/P:1, a s!bstance #e ot to kno# 3revio!sly as an
intermediate of lycolysis.
,4
C is fo!nd in the carboxyl ro!3 of 3/P:. At first
#as it ass!med that the molec!le acce3tin the carbon dioxide #o!ld have to be a
C
8
!nit. &!t after a f!tile search #as finally rib!lose di3hos3hate +R!DP1, a C
-

!nit identified as the acce3tor
C
-
@ C
,
J 8 C
3

This reaction is cataly=ed by rib!lose bis3hos3hate carboxylase +also called
R!bisco or, formerly, fraction/,/3rotein1, as far as ?!antity is concerned the most
common 3rotein of the #orld. The 3rotein com3lex of reen 3lants consists of
eiht times t#o s!b!nits, eiht lare and eiht small ones The 3ict!re to the riht
sho#s 3art of the en=yme toether #ith rib!lose 3hos3hate , C)
8
, and a
manesi!m ion +reen ball1 essential for the reaction. An interactive file
demonstrates the sinle, s!bse?!ent reactions.
After loner reaction 3eriods +- seconds, ,7 seconds, etc.1 #ere f!rther labelled
com3o!nds fo!nd. CA"K4* and &(*2)* determined the se?!ence of the
incor3oration and #ere able to !nite the sinle ste3s to a 3ath#ay. T#o res!lts
#ere es3ecially interestinC
o the resynthesis of rib!lose di3hos3hate and
o the 3rod!ction of the assimilate +the net 3rod!ct of the carbon dioxide
assimilation1.
The 3rod!ction of rib!lose di3hos3hate is best described by a cycle +the CA"K4*
cycle1, #hile the assimilate 3rod!ction is a linear 3rocess. 4t is based on the fact
that an intermediate of the CA"K4* cycle is ded!cted from it.
8. 3/3hos3holycerate is red!ced to lycerinaldehyde/3/3hos3hate +:AP1, the
carboxyl ro!3 is transformed into an aldehyde ro!3. The reaction cons!mes
ATP and *ADP'
8
. The reverse reaction occ!rs, too, in lycolysis tho!h in
3hotosynthesis *ADP is needed instead of the *AD cons!med d!rin lycolysis.
4t is kno#n today that the t#o reactions +and all others, too1 are cataly=ed by
different en=ymes and that the en=ymes of 3hotosynthesis !se *ADP +A
*ADP'
8
1 as a cofactor.
The CA"K4* cycle has to be 3assed three times in order to 3rod!ce one molec!le
of lycerinaldehyde/3/3hos3hate +a CL2;&L3L s!bA !nit1 via 3hotosynthesis
since M!st one molec!le of carbon dioxide is fixed in every ro!nd.
3. %!st as in lycolysis is 3art of the lycerinaldehyde/3/3hos3hate converted into
dihydroxyacetone3hos3hate +DAP1 by e3imeri=ation.
4. Br!ctose/,,0/di3hos3hate +B/,,0/P1 is formed by addition of one molec!le
lycerinaldehyde/3/3hos3hate and one molec!le dihydroxyacetone3hos3hate.
-. Br!ctose/,,0/di3hos3hate is converted into fr!ctose/0/3hos3hate +B/0/P1 by
s3littin off P
i
. The B/0/P has t#o alternative fatesC
o )ne of the B/0/P molec!les is converted into l!cose/0/3hos3hate +:/0/P1
that is sl!iced a#ay from the CA"K4* cycle and is the net yield of
3hotosynthesis.
o The other B/0/P disinterates into a C
-
!nit +xyl!lose/-/3hos3hateN I/-/P1
and a C
,
!nit that forms a C
4
!nit +erythrose/4/3hos3hateN (/4/P1 toether
#ith :AP.
0. The (/4/P is co!3led to one molec!le of dihydroxyacetone3hos3hate +DAP1. The
res!lt is a molec!le of sedohe3t!lose/,,./di3hos3hate +2DP1, a C
.
!nit.
.. After s3littin off one of the t#o 3hos3hate resid!es reacts the sedohe3t!lose/./
3hos3hate +2/./P1 #ith lycerinaldehyde3hos3hate. T#o C
-
!nits are the res!ltC
rib!lose/-/3hos3hate +R!/-/P1 and xyl!lose/-/3hos3hate +I/-/P1.
6. Rib!lose/-/3hos3hate is 3hos3horylated to rib!lose/,,-/di3hos3hate and starts a
ne# ro!nd of the CA"K4* cycle.
4n s!mmary, one can describe the end res!lt of the dark reactions as follo#sC
0 R!DP @ 0 C)
8
A ,8 3/P:
,8 3/P: @ ,8 *ADP'
8
@ ,8 ATP A ,8 :AP @ ,8 ADP @ ,8 P
i
@ ,8 *ADP
,8 :AP A , l!cose +net synthesis 3rod!ct of carbon dioxide assimilation1 @ ,7 :AP
,7 :AP @ 0 ATP A 0 R!DP
*ADP'
8
and ATP stem, as #e #ill see, from the liht reactions of 3hotosynthesis in
#hich the liht enery is converted into chemical enery.
After !nderstandin the 3ath#ay in Chlorella pyrenoidosa arose the ?!estion #hether it
occ!rs in all other reen 3lants, too. 4t co!ld be sho#n to be an im3ortant 3ath#ay of all
reen 3lants. (ven isolated chloro3lasts +from s3inach, for exam3le1 are still f!lly active
and all reactions of the CA"K4* cycle take 3art #ithin them.
C

, C
!
and CA". Re#$lation of The Acti%ity of Photosynthesis
C
!
A n!mber of 3lants dis3lay an increased and more efficient net 3hotosynthesis d!rin
stron liht intensities. A 3rime exam3le are the :ramineae of #armer reions like mai=e
or s!ar/cane.
At the beinnin of the sixties observed '. F)RT2C'AF +'a#aiian 2!ar Planter$s
Association1 that the first 3rod!ct of 3hotosynthesis in s!ar/cane is not the C
3
!nit 3/
3hos3holycerate b!t a !nit #ith fo!r C/atoms. The A!stralian 3lant 3hysioloist M. D.
'ATC' and his (nlish collea!e C. R. 2"ACF confirmed this res!lt and identified the
com3o!nd as oxaloacetate +)AA1. 4t is 3rod!ced by the addition of one molec!le of
carbon dioxide to 3hos3hoenol3yr!vate +P(P1. The cycle is also kno#n as the 'ATC'/
2"ACF/cycle or the C
4
cycle. Plants #ith this cycle are called C
4
/3lants +and CAM
3lants, res3ectively1 in contrast to C
3
3lants #here the carbon dioxide is directly fed into
the CA"K4* cycle. The oxaloacetate is !s!ally converted into malate of #hich the
carbon dioxide is s3lit off aain #ith the hel3 of an en=yme.
This carbon dioxide is no# bo!nd by rib!lose/,,-/di3hos3hate and assimilated via the
CA"K4* cycle. C
2ome s3ecies !se malate instead of as3artate
oxaloacetate @ "/l!tamate A as3artate @ alpha/ketol!tarate.
The reversible bindin of carbon dioxide has the f!nction to acc!m!late and store C)
8
.
The 3rocess cons!mes enery, so that it co!ld also be s3oken of a carbon dioxide 3!m3.
4t sho!ld be mentioned that the 'ATC'/2"ACF cycle re?!ires t#o molec!les of ATP
are 3er fixed carbon dioxide.
Photosynthesis of C
4
plants. C)
8
is bo!nd to 3hos3hoenol3yr!vate +P(P1 in meso3hyll
cells. The 3rod!ct is oxaloacetate. The next ste3 enerates malate. 4n the cells of the
vasc!lar b!ndle sheath, the $Fran=$ cells, is carbon dioxide s3lit off the malate and fed
into the CA"K4* cycle. The 3yr!vate is trans3orted back into the meso3hyll cells +active
trans3ort1 and is #ith the hel3 of additional ATP 3hos3horylated to P(P.
The anatomy of C
4
leaves #ith so/called $Fran=$ cells differs f!ndamentally from that of
C
3
3lants. The chloro3lasts of C
3
3lants are of homoeneo!s str!ct!re, #hile t#o ty3es of
chloro3lasts occ!r in C
4
3lants. The meso3hyll cells contain normal chloro3lasts, that of
the vasc!lar b!ndle sheath have chloro3lasts #itho!t rana , i.e. they are 3artially
im3aired in f!nction. This 3ec!liarity does not affect the CA"K4* cycle, it concerns only
the liht reactions of 3hotosynthesis. The first bindin of carbon dioxide +the 'ATC'/
2"ACF reaction1 occ!rs in the meso3hyll cells, the incor3oration into carbohydrates +the
CA"K4* cycle1 in the cells of the vasc!lar b!ndle sheath. &oth 3rocesses of
3hotosynthesis are s3atially se3arated.
The Crass$lacean Acid "etabolism &CA"'
CAM is the abbreviation of Crass!lacean acid metabolism. The name 3oints at the fact
that this 3ath#ay occ!rs mainly in Crass!lacean s3ecies +and other s!cc!lent 3lants1. The
chemical reaction of the carbon dioxide acc!m!lation is similar to that of C
4
3lants b!t
here are carbon dioxide fixation and its assimilation not se3arated s3atially b!t in time.
CAM 3lants occ!r mainly in arid reions. The o3enin of the stomata to take !3 carbon
dioxide is al#ays connected #ith lare losses of #ater. To inhibit this loss d!rin intense
s!n +the trans3iration via the c!ticle remains intact1 has a mechanism develo3ed that
allo#s the !3take of carbon dioxide d!rin the niht. The 3refixed carbon dioxide is
stored in the vac!oles as malate +and isocitrate1 and is !sed d!rin the daytime for
3hotosynthesis.
(hich "etabolism )oes (ith (hich
Conditions*
The en=yme that cataly=es the 3rimary carbon dioxide
fixation of C
4
and CAM 3lants is 3hos3hoenol3yr!vate
carboxylase +P(PC1. 4ts affinity for carbon dioxide is by far
hiher than that of R!bisco, the first en=yme of the CA"K4*
cycle. As a conse?!ence are C
4
3lants able to !se even trace
amo!nts of carbon dioxide. P(PC occ!rs in small amo!nts
+ro!hly 8 / 3 O1 also in C
3
3lants, #here it, too, has a key
3osition in the metabolic re!lation.
Carbon dioxide yield of C
4
and C
3
plants of open
grasslands in different parts of the world. 4n tem3erate
reions is the rather lo# liht intensity decisive for the
disadvantae of C
4
3lants. C
3
3lants have an advantae d!e
to their lo# rate of 3hotores3iration and beca!se they need
no enery for the 3revio!s fixation of C)
8
. +%. R.
('R"4C'(R, ,9.61.
4n ro#in roots +of mai=e seedlins, for exam3le1 does P(PC hel3 to s!33ly the li3id
synthesis #ith *AD' @ '
@
. The follo#in reactions take 3laceC
,. 3hos3hoenol3yr!vate @ 'C)
3
/ A oxaloacetate @ P
i

8. oxaloacetate A malate. / D!rin this ste3 is *AD' @ '
@
oxidi=ed to *AD
@
.
3. malate A 3yr!vate @ C)
8
.
D!rin the last reaction is *AD red!ced to *AD' @ '
@
.
4n the root nod!les of le!minosae is nitroen fixed. (no!h carbon bodies have to be
s!33lied in order to incor3orate the ammonia 3rod!ced by the bacteria. the C)
8
/bindin
via P(PC/reaction th!s is an im3ortant s!33lement
B!rthermore 3rod!ces the P(PC intermediates of the citric acid cycle +oxaloacetate and P
or malate1, a back/!3 in case of shortaes. The activity of P(PC is controlled by extern
factors, the day lenth is decisive. 4n some cases have different isoen=ymes be fo!nd in
different tiss!es. Their 3rod!ction is controlled by different triers.
C
3
3lants can loose !3 to 87 3ercent of the carbon fixed in the CA"K4* cycle at intense
radiation. ;nder stron liht is the 3hotores3iration ,.- / 3.- times as hih as the !s!al
res3iration in darkness. 4n C
4
3lants becomes the 3hotores3iration drastically red!ced, it
may even not be detectable any more. 4n other #ordsC
The net rate of 3hotosynthesis +and conse?!ently also the net 3rod!ction of biomass1 of
C
4
3lants is far larer at hih liht intensities than that of C
3
3lants. The o3timal
tem3erat!re of 3hotosynthesis is belo# that of the res3iration in darkness. As a
conse?!ence are losses ca!sed by res3iration larer at hih than at lo# tem3erat!res.
>here liht is a limitin factor and tem3erat!res are lo# +i.e. in tem3erate climatic
=ones1 have C
3
3lants the advantae, C
4
3lant do hardly occ!r +one of the exce3tions is
Spartina townsendii1. C
4
3lants, nearly al#ays herbs or shr!bs, are more s!ccessf!l in the
o3en co!ntry of #armer =ones.
4t has to be mentioned that t#o molec!les of ATP are cons!med in the 'ATC'/2"ACF
cycle
C
4
3lants belon to n!mero!s, 3hyloenetically not related monocotyledono!s and
dicotyledono!s families. Moreover have C
4
activities also been detected in the bl!e/reen
ala Anacystis nidulans as #ell as in some dinoflaellates.
2ince the alternative C
3
or C
4
is accom3anied by considerable chanes of the leaf
anatomy has it to be ass!med that the enetic 3otential for both 3ath#ays is ?!ite
common in the 3lant kindom and that, de3endin on the ecoloical needs, one #ay is
chosen by a s3ecies #hile a related s3ecies may choose the other one.
A #ell/st!died exam3le is the en!s Atriplex, #here both #ays are reali=ed. The C
3

3lants belon to one 3hyloenetic ro!3, the C
4
3lants to another. 4n some cases can
hybrids of C
3
and C
4
s3ecies be enerated.
4nfl!ence of different 3arameters on the efficiency of the carbon dioxide !3take
+ordinate1 of a C
3
3lant +Atriplex patula, yello# line1 and a C
4
3lant +Atriplex rosea, reen
line1. Meas!red 3arameters +from left to riht1C liht intensity, leaf tem3erat!re and
concentration of carbon dioxide #ithin the intercell!lar s3ace +accordin to ).
&%QRFMA* and %. &(RR5, ,9.31.
4n several 3lant s3ecies of the enera Zea, Mollugo, Moricandia, Flaveria, etc. occ!r both
ty3es of C)
8
fixation #ithin one 3lant. 4n yo!ner 3lants is !s!ally the C
3
/, in older ones
the C
4
3ath#ay taken. The amo!nt of C
4
is controlled by environmental factors.
CAM: Advantages and Disadvantages. CAM has been detected in more than ,777
anios3erms of ,. different families. 4t is !s!ally accom3anied by s!cc!lence, tho!h not
all Crass!laceae, for exam3le, dis3lay CAM and s!cc!lence is no 3recondition of CAM.
Tillandsia usneoides of the bromelia family is not s!cc!lent, b!t !ses CAM.
Mesemryanthemum crystallinum +a 3lant #ith s!cc!lent leaves1 can !se the C
3
3ath#ay
b!t s#itches to CAM #hen ro#in in saline soils. ;nder ex3erimental conditions can
the shift be achieved by increasin the *aCl concentration of the n!trient medi!m +F.
>4*T(R and D. %. von >4""(RT, ,9.81. >hile the advantae of C
4
3lants comes in
!sef!l !nder hih liht intensities, is the deree of the CAM infl!ence in CAM 3lants
re!lated mainly by tem3erat!re, atmos3heric h!midity and salinity. &oth stron and
#eak CAM 3lants are kno#n. 4n #eak CAM 3lants becomes CAM only a33arent at
certain differences bet#een day and niht tem3erat!re. CAM 3lants that store a lot of
malate and d!e to the th!s hih osmotic val!e also a lot of #ater, are !s!ally less frost
resistant than C
3
3lants. &eca!se of the hih concentration of acid are they less heat
resistant, too. 23ecies of arid reions are therefore forced to break their 3ool of malate
do#n d!rin the daytime +R. "Q2C' and '. FAPP(*, ;niversitRt Fiel, ,96-1. ;s!ally
do the C
4
3ath#ay and CAM excl!de each other. An exce3tion is the s!cc!lent C
4

dicotyledon ortulaca oleracea that is able to choose the o3timal 3ath#ay. !nder nat!ral
conditions
The Li#ht Reactions of Photosynthesis
4t has been mentioned in the historical o!tline that 3hotosynthesis is de3endent on liht.
The res!lts of (*:("MA** and 2AC'2 sho#ed that it is absorbed by chloro3hyll. >e
also ot to kno# that 3lants have t#o ty3es of chloro3hyll, chloro3hyll a and b and that
both ty3es dis3lay a characteristic absor3tion s3ectr!m.
The action s3ectr!m of 3hotosynthesis resembles the absor3tion s3ectra of chloro3hyll
tho!h it is not identical. This means that f!rther 3hotorece3tors +so/called accessory
3iments1 exist.
>e already ot to kno# somethin abo!t the assimilation of carbon dioxide in the
3revio!s section. )!r ?!estion is no#C #hich reactions are ind!ced by liht and ho# is
the liht enery converted into chemical eneryS )r, in other #ords, ho# are ATP and
*ADP'
8
3rod!cedS
CA"K4* and his collaborators st!died the dark reactions in intact, active cells. This
attem3t 3roved to be ins!fficient for the liht reactions. The res!lts #ere contradictory.
Techni?!es to isolate active chloro3lasts had to be develo3ed.
After the !se of fractions containin isolated chloro3lasts became !s!al, #ere three
research ro!3s at the same time +,9-,1 and inde3endent of each other able to sho# that
isolated chloro3lasts red!ce *ADP to *ADP'
8
#hen ex3osed to liht G>. K42'*4AC
and 2. )C')A +Rockefeller 4nstit!te, *e# 5ork1, ". %. T)"MAC' +;niversity of
Chicao1 and D. 4. AR*)* +;niversity of California, &erkeley1H. 2hortly after#ards +in
,9-41 discovered AR*)* and his collaborators that the 3rod!ction of ATP, too, is
de3endent on liht and that both ATP and *ADP'
8
can be 3rod!ced sim!ltaneo!sly.
&oth com3o!nds are enerated from 3rec!rsors that are 3resent in the chloro3last already
before 3hotosynthesis starts, since no extern metabolites #ere s!33lied d!rin the
ex3eriments. Accordinly #as liht +3hotons1 the only available so!rce of enery. 4t
t!rned o!t that the 3rod!ction of ATP needs no oxyen, neither is oxyen 3rod!ced
d!rin the reaction. Conse?!ently r!ns the e?!ation as follo#sC
n ADP @ n P
i
A (photons) A n ATP
The 3rocess is termed photophosphorylation. 4t exists in bacteria and bl!e/reen alae,
too, and is a eneral feat!re of 3hotosynthetic 3rocesses.
After it #as 3roven hat ATP is 3rod!ced, #as it asked ho# this is done. 4t seemed
!nlikely that the liht ind!ces the 3rod!ction of ATP directly b!t it had t!rned o!t that
the 3rod!ction of ATP has to be 3receded by an ex3os!re to liht. The conce3t of a liht/
ind!ced electron flo# #as develo3ed. 4t ass!mes that one molec!le of chloro3hyll
absorbs one 3hoton. As a conse?!ence is an electrons of chloro3hyll transferred to a
hiher enery level.
This enery/rich electron is then transferred to a neihbo!rin electron acce3tor #ith a
stron electroneative redox 3otential. The transfer of the electron from the activated
chloro3hyll to the +first1 acce3tor is the first 3hotochemical 3hase of 3hotosynthesis. 4ts
decisive feat!re is the transformation of a 3hoton flo# +liht1 into a flo# of electrons.
As soon as a stronly electroneative +red!cin1 s!bstance has been 3rod!ced can the
electron flo# 3roceed #ith electron acce3tors of less neative redox 3otentials. The
3rocess releases chemical enery that is !sed for 3hoto3hos3horylation. Already d!rin
the fifties existed the first stron 3roves for the involvement of the chloro3lasts$
cytochromes. 4t co!ld be sho#n, too, that the electron is finally acce3ted by a
chloro3hyll, so that its oriinal state is restored aain. The re?!irements of catalysis are
f!lfilled. The 3rocess became kno#n as cyclic phosphorylation +D. 4. AR*)*, ,9-91.
2!ch a cyclic flo# of electrons that is 3o#ered by liht and releases chemical enery
!sed for the 3rod!ction of ATP is !ni?!e. 4t is the o!tstandin 3ro3erty of 3hotosynthetic
cells.
Concept of cyclic photophosphorylation +to the left1. To the rihtC the oriinal conce3t
of non/cyclic 3hoto3hos3horylation +accordin to D. 4. AR*)*, ,9.,1
The only !nex3lained 3rocess remained #as the 3hotored!ction of *ADP in chloro3lasts.
4t #ere aain AR*)* and his collaborators that #ere in ,9-. able to discover a second
3art of 3hoto3hos3horylation. They co!ld 3rove ex3erimentally that the 3hotored!ction
of *ADP and the synthesis of ATP are co!3led. 4n contrast to the cyclic
3hoto3hos3horylation is the 3rod!ction of ATP co!3led stoichiometric to a liht/ind!ced
transfer of electrons from #ater to *ADP and to the 3rod!ction of oxyen. The ATP
3rod!ction of the #hole system increases the red!ction rate of *ADP. This 3ointed at the
fact that the 3rocess is tihtly co!3led to cyclic 3hoto3hos3horylation. 2ince electrons are
irreversibly transferred from chloro3hyll to *ADP, are s!bstit!tes needed and these
electrons stem from the breakdo#n of #ater. 4t is s3oken of non-cyclic
photophosphorylation, since ATP is 3rod!ced sim!ltaneo!sly. Berredoxin +a heme/less
iron/s!l3h!r 3rotein1 has a key 3osition in this 3rocess. 4ts red!ction 3otential is far more
neative than that of *ADP so that an electron flo# from ferredoxin to *ADP #as very
likely. The red!ction of *ADP is a three ste3 reactionC
,. a 3hotochemical red!ction of ferredoxin that is follo#ed by t#o $dark$ ste3s.
8. The re/oxidation of ferredoxin #ith the hel3 of a ferredoxin/*ADP/red!ctase +a
flavo3rotein1.
3. The re/oxidation of the ferredoxin/*ADP/red!ctase by *ADP.

erredoxin: 4ron/s!l3h!r/com3lexC / to the leftC Be
8
2
8
3rotein / Plantty3e ferredoxins , to
the rihtC Be
4
2
4
3roteins / &acterialty3e ferredoxins
fromC PR)M42( / The Prosthetic ro!3s and Metal 4ons in Protein Active 2ites Database
>hat #as at first rearded as a 3hotored!ction of *ADP 3roved to be an electron
trans3ort chain that r!ns from ferredoxin via a flavin com3onent to *ADP.
The o!tstandin 3osition of ferredoxin #as strenthened even more after it #as fo!nd o!t
that stoichiometric amo!nts of )
8
and ATP are 3rod!ced d!rin the reaction. The non/
cyclic 3hoto3hos3horylation can accordinly be described as follo#sC
4 ferredoxin +oxidi=ed1 @ 8 ADP @ 8 P
i
@ 8 '
8
) A (photons) A 4 ferredoxin +red!ced1 @ 8
ATP @ )
8
@ 4 '
@
.
The res!lts led to the ?!estion ho# this reaction is co!3led to the cyclic
3hoto3hos3horylation disc!ssed at the beinnin. A series of ex3eriments !sin s3ecific
inhibitors sho#ed that ferredoxin is a com3onent of that 3ath#ay, too.
2o m!ch abo!t the chemical data. More kno#lede abo!t the 3rimary effect of the liht
and abo!t the sinificance of chloro3hyll #o!ld have to exist to inter3ret them. These
3roblems, too, have a lon 3ast history.
T+o Photosystems
4n ,938 ex3osed R. (M(R2)* and ;. AR*)"D of the ;niversity of 4llinois at ;rbana
Chlorella cells to a series of extremely short flashes of liht. >ith this ex3eriment did
they try to find o!t ho# many molec!les of chloro3hyll #ere necessary to !se one 3hoton
for the 3rod!ction of one molec!le of oxyen. The res!lt #as that several h!ndred
chloro3hyll molec!les are necessary #hich means that not all of them are of the same
im3ortance. Most act as liht tra3s +or antennas1 hel3in to transfer a 3hoton to a reaction
centre #here an es3ecially ex3osed chloro3hyll transforms liht enery into chemical
enery. '. :ABBR)* called this com3lex of several h!ndred chloro3hyll molec!les and
other 3iments +carotenes, carotenoids, xantho3hylls, etc.1 a photosynthetic !nit.
This areation of 3iments seems to lead to an es3ecially efficient !se of the incomin
liht. 2till, a rather lare 3art of the irradiated enery is lost. 4t does never reach the
reaction centre and is emitted as #armth or liht +red a!tofl!orescence of chloro3hyll1.
>hen reardin the absor3tion s3ectr!m of 3hotosynthesis does it stand o!t that the
efficiency of liht of the #ave lenth lam!da A 067 nm decreases stronly altho!h
chloro3hyll a dis3lays an absor3tion in that area. R. (M(R2)* +,9-.1 discovered that
liht of the #ave lenth lam!da A .77 +.,71 increases the rate of 3hotosynthesis
drastically if liht of the #ave lenth lam!da J 067 nm +or less1 is 3resent at the same
time.
>hen these t#o liht ?!alities are !sed inde3endent of each other or one after the other is
no increase meas!red. (M(R2)* concl!ded that t#o 3hotochemical 3rocesses have to
exist that consist of different 3iment systems +liht rece3tors1 b!t that do co/o3erate
+(M(R2)*/effect1. Accordin to a s!estion of ". *.M. D;52(*2 are the t#o
systems called
photosyste" # $P% #&. 4t needs liht of loner #ave lenths +lam!da A .77 nm1
and
photosyste" ## $P% ##&. 4t becomes active #hen ex3osed to shorter #ave lenths
+lam!da L 067 nm1
The ratio of chloro3hyll a to chloro3hyll b is hiher in P2 4 than in P2 44. The ?!estion
ho# the t#o systems co/o3erate and ho# they are co!3led to the 3rod!ction of ATP and
*ADP'
8
remains to be settled.
AR*)* and his collaborators co!ld 3rove that the t#o systems are arraned in series and
that both systems are re?!ired to ex3lain all effects that had been reconi=ed as
3hotosynthetic ones. )nly some bacteria that 3rod!ce no oxyen d!rin 3hotosynthesis
lack 3hotosystem 44. This res!lts hints at the s!estion that the s3littin of #ater is
co!3led to 3hotosystem 44 and that 3hotosystem 4 develo3ed earlier in evol!tion.
The reaction centre of every 3hotosystem is re3resented by one molec!le of chloro3hyll a
each +P .77 in P2 4 and P 067 in P2 44, #here P means 3iment1.
The absor3tion of a 3hoton by P 067 +#hich has a 3ositive redox 3otential of @ 7,6 K in
its basic state1 transfers P 067 into its excited state + #ith a redox 3otential of 7,7 K1 and
ca!ses the formation of a stronly oxidi=in com3onent +<
@
1 and a #eakly red!cin +T
/
1
one. <
@
#ithdra#s electrons from #ater so that )
8
and 3rotons are set free.
4 <
@
@ 8 '
8
) A 4 < @ 4 '
@
@ )
8

The red!cin com3onent +a membrane/bo!nd 3lasto?!inone1 feeds the electron into an
electron trans3ort chain in the co!rse of #hich it looses enery 3art of #hich is !sed for
the 3rod!ction of ATP. The electron does not ret!rn to its startin 3oint +chloro3hyll P
0671 b!t is transferred to a chloro3hyll molec!le of 3hotosystem 4 +P .771. The t#o
3hotosystems are th!s co!3led.
The absor3tion of a f!rther 3hoton excites the P .77 M!st mentioned +the redox 3otential
of #hich is @ 7,4 / @ 7,- in its basic state1. 4t transfers one electron to a membrane bo!nd
ferredoxin +P4371 #hich 3asses the electron on to a sol!ble ferredoxin. The follo#in
ste3s are kno#n.
4n a stoichiometric sense is the o!tline above incom3lete since ferredoxin transfers M!st
one electron at any iven time #hile t#o electrons are needed for the 3rod!ction of one
*ADP'
8
. The e?!ation #o!ld conse?!ently have to beC
8 ferredoxin +red!ced1 @ 8 '
@
@ *ADP
@
A 8 ferredoxin +oxidi=ed1 @ *ADP'
8

4n s!mmary is the liht enery !sed for the flo# of electrons from #ater to *ADP'
8
and
for the sim!ltaneo!s 3rod!ction of ATP +</scheme1.
D!rin o!r disc!ssion did #e nelect the cyclic 3hoto3hos3horylation mentioned at the
beinnin. 4t 3roved to be a 3arallel 3rocess that starts #ork as soon as eno!h *ADP'
8

b!t too small amo!nts of ATP are 3resent. )nly 3hotosystem 4 3artici3ates in cyclic
3hoto3hos3horylation.
The Photosynthetic "embrane
4n o!r disc!ssion of 3hotosynthesis have #e th!s far only rearded biochemical reactions.
4n the section abo!t lycolysis and other biosynthetic 3ath#ays #as the sinificance of
sinle en=ymes 3ointed o!t. 2ince ?!ite some time kno#, for exam3le, have all en=ymes
involved in lycolysis been 3!rified and isolated and each ste3 of the 3ath#ay can be
analy=ed !nder in vitro conditions. 4t has also been tried to isolate the com3lete set of
com3onents necessary for 3hotosynthesis in order to reconstit!te the #hole system. &!t
all attem3ts failed beca!se the 3remises they #ere based on #ere #ron as #e kno#
today.
A n!mber of 3roblems have not been taken into consideration !ntil no#. The terms
3hotosystem 4 and 3hotosystem 44, for exam3le, have been introd!ced and all
3artici3atin 3iments have been mentioned b!t the follo#in s!bMects remain to be
disc!ssedC
'o# are the 3hotosystems orani=edS
'o# are the 3iments arranedS
>hy does one of the chloro3hyll molec!les react different than all the othersS
>hy are action and absor3tion s3ectra not ?!ite conr!entS
>hy reacts P 067 +chloro3hyll a1 different than P .77 +chloro3hyll a, too1S
'o# are electron trans3ort chain and ATP 3rod!ction co!3ledS
'o# are 3hotosystem 4 and 44 linkedS
>hich str!ct!ral 3rere?!isites have to exist in order for the t#o systems to co/o3erateS
4t #as al#ays acce3ted that each of the biochemical reactions #as cataly=ed by a s3ecific
en=yme and still, it took ?!ite some time before it #as reali=ed that the chloro3hyll and
the other 3iments are 3rotein/bo!nd and that they are only active as 3rotein/chloro3hyll
+and 3rotein/3iment, res3ectively1 com3lexes. The isolated 3iments themselves #ere
!seless for 3hotosynthesis. The 3iment/3rotein com3lex, +most1 3roteins of the electron
trans3ort chain as #ell as the catalyst of ATP synthesis +ATP synthase1 are interal
com3o!nds of the 3hotosynthesis membrane+s1 +J the thylacoid membranes of alae and
hiher reen 3lants, cyto3lasmatic membranes of 3hotosynthetically active bacteria and
bl!e/reen alae1. The location #ithin the membrane +at the o!t/ or the inside, for
exam3le1 and the relative arranement of the 3roteins to#ards each other are im3ortant
3rere?!isites of enery transformation.
This is not only tr!e for 3hotosynthetic reactions b!t also for those of the res3iratory
chain and for the en=ymes located #ithin the 3!r3le membrane of "alo!acterium
halo!ium +an archaebacteri!m !sin liht enery for the 3rod!ction of ATP #itho!t an
electron flo#1.
The re?!irements for enery transformation are even hiherC com3letely intact
membranes that are im3ermeable for 3rotons and that enclose com3artments th!s
maintainin a stable electrochemical radient bet#een inside and o!tside. The 3rod!ction
of ATP is based on a directed 3roton dislocation 3aralleled by a chane of the
com3artment$s 3' and of its membrane 3otential.
Proteins of The Photosynthetic "embrane
The research into the 3roteins essential for 3hotosynthesis started very late. The reason is
that all of them are membrane/bo!nd #hich rendered it nearly im3ossible to isolate and
characteri=e them #ith the classical methods of 3rotein analysis.
)nly after sensitive techni?!es like el electro3horesis and the controlled !se of
deterents like sodi!m dodecyl s!lfate +2D21 had been develo3ed, became it 3ossible to
se3arate the 3roteins and to identify them as bands in a el. A side 3rod!ct of this
techni?!e is the determination of the molec!lar #eihts of the res3ective 3oly3e3tide
chain.
A second, inde3endent attem3t #as and is the !se of s3ecific 3robes like fl!orescence/
taed antibodies that hel3 to find o!t #hether a certain 3rotein +or 3art of a 3oly3e3tide
chain1 is located at the inside or the o!tside of a membrane. The !se of antibodies aainst
s3ecific 3roteins allo#s, too, to 3reci3itate these 3roteins selectively since only they are
able to form the extremely s3ecific antien / antibody com3lex.
Cross/linkin aents render it 3ossible to el!cidate the s!rro!ndin of a molec!le. And
the !se of s3ecific inhibitors hel3s locali=in their site of effect. DCM; G3/+3$, 4$ /
dichlor3henyl1 / ,,, / dimethyl!reaH has since years been !sed to inhibit 3hotosystem 44.
4t has no effect on 3hotosystem 4 and #as therefore !sed by AR*)* and his
collaborators as an im3ortant hel3 to st!dy the electron trans3ort chain that starts at
3hotosystem 4 inde3endently of that ind!ced by 3hotosystem 44.
>e kno# today that DCM; does not effect chloro3hyll itself b!t a certain 3rotein, the
3lasto?!inone/bindin 3rotein.
A third 3ossibility to characteri=e the 3hotosynthetic membrane is the analysis of certain
m!tants. The sinle/celled ala Chlamydomonas reinhardii 3roved to be a ood test
obMect. T!ite a rane of m!tants #ith 3hotosynthetic defects are kno#n. They can be
ro!3ed in fo!r classesC
,. m!tants #ith a defect in 3hotosystem 4,
8. m!tants #ith a defect in 3hotosystem 44,
3. m!tants #ith a defect in 3hoto3hos3horylation and
4. m!tants #ith a defect in the antenna com3lex.
4t is ?!ite strikin that almost all m!tants are characteri=ed not only by the loss or chane
of a certain 3oly3e3tide chain b!t by the lack of a #hole com3lex, for exam3le that of P2
4. 4t seems therefore as if the m!tations #o!ld lead to 3leiotro3ic effects. )r, ex3ressed
differentlyC #hen a 3oly3e3tide chain is chaned or missin does the assembly of the
other 3oly3e3tide chains not #ork any more. This observation sho#s ho# tiht the
interactions bet#een the sinle 3oly3e3tide chains are and ho# im3ortant they are for
their m!t!al co/o3eration.
A f!rther and not less im3ortant techni?!e is electron microsco3y !s!ally !sed in
combination #ith free=e/etchin.
The se?!encin of membrane 3roteins remains diffic!lt. And yet, the se?!ences of most
3roteins involved in 3hotosynthesis co!ld be determined d!rin the last years via the
se?!encin of their res3ective enes. The most remarkable o!tcome of this #ork is that
these 3roteins contain +M!st like 3roteins of animal or bacterial membranes, too1 a lare
3ortion of alpha / helices. The lenths of the helices corres3onds to the thickness of the
membranes. The sinle helices are connected via 3olar and P or non/hydro3hobic
se?!ences.
Chloro,hyll - .indin# Proteins
2everal chloro3hyll/bindin 3roteins of the 3hotosynthetic membranes of different
systematic ro!3s +anios3erms, ymnos3erms, alae, bacteria1 have been isolated and
characteri=ed. The best/kno#n are P .77/chloro3hyll/a/3rotein , and the liht/harvestin
chloro3hyll/aPb/3rotein 8 #hich have been st!died in the laboratory of %. P.
T')R*&(R:(R at the ;niversity of California, "os Aneles. &oth are stronly
hydro3hobic, interal membrane 3roteins. &oth bind chloro3hyll a b!t only the latter
binds chloro3hyll b, too. The P .77/chloro3hyll/a/3rotein , contains the reaction centre
+P .771 of 3hotosystem 4, i.e. one of the chloro3hyll molec!les is bo!nd in a s3ecific
confi!ration and is located in a s!rro!ndin +d!e to a s3ecific amino acid com3osition
and the foldin of the 3oly3e3tide chain1 that renders it different than all other
chloro3hyll molec!les bo!nd by this 3rotein, too. This str!ct!ral 3ec!liarity is the
3recondition for the liht/ind!ced activation and conse?!ently for the ind!ction of the
electron flo#.
The molec!lar #eiht of the 3oly3e3tide chain is ,,7,777 Dalton. 4t is able to bind ,4
chloro3hyll molec!les. The liht/harvestin/3rotein +liht/harvestin chloro3hyll/aPb/
3rotein 81 is also very common. 4t is mainly associated #ith 3hotosystem 44 b!t effects on
3hotosystem 4 have been observed, too. Chloro3hyll a and b are bo!nd in e?!imolar
amo!nts besides l!tein and !eta / carotene. The chloro3hyll to carotenoid ratio is 3 / . C ,
on a molec!lar level.
Structure of the light harvesting complex of photosystem II -
Arrangement of Pigments
# Copyright $%%&, Antony Cro'ts, (niversity o' )llinois at (r!ana*Champaign, a*cro'ts+uiuc,edu
Light Harvesting Complexes I and II (LH I, LH II and !eaction Centre
(!C
UC Theoretical &ioloy :ro!3 / ;niversity of 4llinois at ;rbana/Cham3ain
The Co$,lin# /actor0 an ATP - 1ynthase
ATP / synthase is the en=yme that cataly=es the synthesis of ATP. 2ince the 3rod!ction of ATP occ!rs not
only d!rin 3hotosynthesis b!t d!rin res3iration, too, s!ested the idea that the ATP 3rod!ction of both
cases is based on similar mechanisms itself.
After havin collected ex3erience #ith the mitochondrial ATP synthase, did (. RACF(R of the Cornell
;niversity isolate an en=yme of thylacoid membranes that resembled the res3ective mitochondrial en=yme
very m!ch. 4n the electron microsco3e did it look like a stemmed knob. The knob #as termed CB, and the
stem CB7 +B, and B7 res3ectively in the mitochondrial en=yme1. CB7 +or B71 is a membrane anchor. >hile
the B, / B7 com3lex is locali=ed in the inner mitochondrial membrane and the knob is directed to#ards the
mitochondrial matrix are the res3ective molec!lar 3arts of the CB7 / CB, com3lex fo!nd at the o!tside of
the thylacoid membranes. 4n both cases consists the ATP synthase o!t of several different 3oly3e3tide
chains, it is an en=yme com3lex. The 3hos3horylation of ADP #orks only, if the ATP synthase is a
com3onent of an intact, 3roton/3ermeable membrane. 4t has to se3arate t#o com3artments +the inner 3art of
the vesicle and the s!rro!ndin1.
#Copyright $%%&, Antony Cro'ts, (niversity o' )llinois at (r!ana*Champaign, a*cro'ts+uiuc,edu
The Chemiosmotic 2y,othesis of P. "ITC23LL0 a "odel of The
Photosynthetic "embrane
2ince ?!ite some time has the ATP synthase been ascribed the f!nction of a co!3lin factor. This means
that it is able to !tili=e the free enery released by electron trans3ort. 2!ch enery conservation is referred
to as energy co!pling or energy transd!ction.
'o# does this #orkS 4t miht have been ass!med that the electron trans3ort chain serves the 3rod!ction of
enery/rich intermediates and that these constit!te an enery store for the 3rod!ction of ATP. T#o
ar!ments aainst this idea existC
,. *o s!ch s!bstances have ever been isolated and
8. 3hoto3hos3horylation +and the oxidative 3hos3horylation of the res3iratory chain, res3ectively1 is
only 3ossible if the thylacoid membranes +the inner mitochondrial membrane, res3ectively1 are
intact.
Another ass!m3tion had been that the ATP synthase chanes its confi!ration and is th!s itself transferred
into an activated state. 4n s!ch a case #o!ld the enery be transiently stored in #eak interactions. This
hy3othesis, too, failed to #ithstand ex3erimental scr!tiny.
4n ,90, 3ro3osed P. M4TC'("" +:lynn Research "aboratories, :reat &ritain1 that the enery set free
d!rin the electron trans3ort is conserved as a 3roton radient across the membrane. The enery #o!ld then
not be stored as a chemical bond b!t as an electrochemical radient. The electrochemical 3otential of this
radient #o!ld be harnessed to synthesi=e ATP. The hy3othesis ex3lains several key observationsC
,. 4t is consistent #ith the fact that oxidative 3hos3horylation re?!ires an intact inner mitochondrial
membrane.
8. The inner membrane is im3ermeable to ions like '
@
or )'
/
#hose free diff!sion #o!ld dischare
the electrochemical radient.
3. The electron trans3ort res!lts in the trans3ort o!t of intact mitochondria +o!t of the thylacoid s3ace
of chloro3lasts1 thereby creatin a meas!rable electrochemical radient across the inner
mitochondrial membrane +the thylacoid membrane of chloro3lasts1.
4. 2!bstances that increase the 3ermeability of the inner mitochondrial or the thylacoid membrane to
3rotons, thereby dissi3atin the electrochemical radient, allo# electron trans3ort to contin!e b!t
inhibit ATP synthesisC they $!nco!3le$ electron trans3ort from oxidative 3hos3horylation.
A very convincin ex3eriment #as 3erformed by A. T. %A:(*D)RB +,900, Cornell ;niversity, 4thaca, *.
5.1C he took isolated thylacoids and inc!bated them in a 3' b!ffer !ntil the same 3' +41 #as meas!red at
both sides of the membrane. After the e?!ilibri!m had been achieved, did he ?!ickly transfer the thylacoids
to a media of 3' 6 containin both ADP and Pi. 4mmediately after the transfer #as the 3' bet#een inside
and o!tside evened o!t #hile, at the same time, the system 3rod!ced ATP. The 3roton flo# across the
membrane #as !sed for the 3rod!ction of ATP. The ex3eriment #orked only #ith intact membranes. 4n
addition have all biochemical 3rocesses to be directed +vectorial1. All en=yme molec!les have to have the
same direction so that the 3rotons are trans3orted in only one direction.
'o# can a flo# of 3rotons ind!ce the 3rod!ction of ATPSS
To !nderstand the 3rocess is it !sef!l to st!dy the ATP 3rod!ction a little more. ADP and 3hos3hate +Pi1
are its startin com3o!nds. 4t is kno#n that both are bo!nd to neihbo!rin b!t se3arate bindin sites of the
en=yme com3lex +the ATP synthase 1. To 3rod!ce ATP +ADPVP1 has one '
@
to be removed from ADP and
one )'
/
from 3hos3hate. 4n a formal sense is #ater s3lit off. As soon as the ions have left the com3lex do
both combine #ith their co!nterions to #ater +not #ith each other1. The end res!lt is a directed flo# of
3rotons.
This does not mean that one 3roton is transferred thro!h the membrane via the ATP synthase com3lex.
4nstead is a ne#ly formed 3roton iven off into sol!tion at one side #hile another 3roton is ca3t!red and
ne!trali=ed +by a )'
/
ion1 at the other side of the membrane.
4nde3endent of these observations co!ld :RW&(R and >4TT +Max/Kollmer/4nstit!t, Technische
;niversitRt &erlin1 sho# that a direct co!3lin bet#een 3roton radient and the electron trans3ort chains of
the 3hotosystems 4 and 44 exists. 4t emered that the already kno#n $<$/scheme is no hy3othetical 3rod!ct
b!t that the involved com3onents are arraned like a < #ithin the 3hotosynthetic membrane, i.e. the
str!ct!ral basis for the 3rocess disc!ssed in the 3revio!s sections became kno#n.
D. von >(TT2T(4* and R. P. )"4K(R +Carlsber "aboratori!m, Co3enhaen, ,96-1 s!mmari=ed all
res!lts and #ere th!s able to develo3 a model that ex3lains the to3oloy of the sinle 3rotein com3lexes.
The 3hotosynthetic membrane contains fo!r com3lexes altoether. (ach has s!b!nits encoded in the
n!cle!s and others encoded by 3lastids. 2everal 3roteins bind chloro3hyll a, one binds chloro3hyll a and b.
The P2 44 com3lex is mainly locali=ed in stacked, P2 4 and the ATP synthase com3lex +CB, / CB71 in non/
stacked thylacoid membranes.
A f!rther remarkC the reactions of the CA"K4* cycle are cataly=ed by sol!ble en=ymes, locali=ed #ithin
the stroma.
"olec$lar 1tr$ct$re of The Reaction Centre
4n ,96- #as the str!ct!re of the 3hotosynthetic reaction centre$s 3rotein s!b!nits determined. %.
D(42(*')B(R, '. M4C'(" and R. ';&(R +Max/Planck/4nstit!t fEr &iochemie, Martinsried1
s!cceeded in crystalli=in the 3rotein com3lex of the 3hotosynthetic membrane of the bacteri!m
-hodopseudomonas viridis, they determined the foldin of the 3oly3e3tide chain and the arranement of
the chloro3hyll molec!les. 4n ,966 #ere they a#arded the *obel 3ri=e for this #ork. Toether #ith the
tertiary and ?!aternary str!ct!re #as also the amino acid se?!ence of the involved 3oly3e3tides
determined. The central 3art of the com3lex contains t#o s!b!nits, " and M, each of #hich forms - helices
that s3an the 3hotosynthetic membrane. T#o f!rther 3oly3e3tides, ' and a cytochrome c / like 3rotein are
associated. B!rthermore belon 4 covalently linked heme ro!3s, 4 bacteriochloro3hyll b molec!les, 8
molec!les of bacterio3heo3hytin b, 8 ?!inones, , iron ion that is not linked to a heme ro!3 as #ell as
carotenoids as 3rosthetic ro!3s to the com3lex. The str!ct!res fo!nd in bacteria are homoloo!s to the
reaction centres of the 3hotosystems 4 and 44 of reen 3lants.