LECTURE 27: PHOTOSYNTHESIS

CHEMISTRY INSIDE THE CHLOROPLAST

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Light reaction, light dependent reaction

Dark rection, light independent reaction

LIGHT REACTION
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Purpose is to generate reducing equivalents in form of nicotinamide, NADP+, converting it into NADPH
o Does so by using water, converting/oxidizing water to molecular oxygen
o Which happens to be a very convenient side product for all of us

DARK REACTION
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NADPH generated by light reaction is used in combination with ATP for CO2 fixation
o Binding CO2 from the atmosphere and converting it into sugars
o The plant does this to build construction material
For cellular walls in the form of cellulose
o But also energy storage
In the form of glucose
Amylopectin in starch, which is the feed for seeds

CHEMISTRY OF THE LIGHT REACTION

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The makeup of the chloroplast, where photosynthesis takes place

o the characteristic feature of the chloroplast is the stacked thylakoid membrane (granum)
o Integral part of thylakoid membranes, embedded in the membrane, are so called PIGMENTS
Light-harvesting pigments (antenna molecules)
Clusters of pigment molecules are embedded in thylakoid membrane
The most important pigment components are these CHLOROPHYLLS
Chlorophylls: small molecules that consist of two parts
1. Porphyrin ring portion: the light-absorbing head of molecule
o Chelates a magnesium
o Responsible for absorbing light energy, the photon that strikes the leaf surface
o Photon is absorbed by porphyrin ring system
o Not just floating around, anchored by:
2. Hydrocarbon tail: anchors porphyrin ring system to membrane,
Best comparison for pigments are water lilies
Water surface is the thylakoid membrane
Individual pigments/chlorophylls are like leaves
Porphyrin ring portion are floating on the membrane surface
Held in place by hydrocarbon tail, like an anchor, locks leaf from floating away
These pigments embedded in thylakoid membrane are harvesting device for energy, photons
That cover a large surface area
The probability that a light particle, a photon, that hits the leaf surface, is likely to hit one of those pigments
What happens when a photon hits one of those perforin ring systems?
Turns out the perforin ring systems have a large, delocalized electron system
Its a conjugated system
As the photon strikes the conjugated electron system, the chromophore (perforin is a chromophore)
Chromophore: part of a molecule responsible for its color. The color arises when a molecule absorbs certain
wavelengths of visible light and transmits or reflects others
One of the electrons in the conjugated system, will absorb the energy delivered by the photon
It will increase the energy of the electron from the ground state to the excited state
The electron doesnt go anywhere, it simply, the energy content of the electron is significantly increased by the energy
delivered from the photon
What do we know about excited states? (Think back to general chemistry)
EXCITED STATE ELECTRONS CANNOT REMAIN IN EXCITED STATE FOR A LONG PERIOD OF TIME
Lifetimes of a fraction of a second
Very, very short-lived
Because the energy that the electron has absorbed, that gets it to the excited state, that energy must be moved on

Turns out excited state electrons have different pathways in which to dissipate this energy:
Those mechanism are shown here on the bottom
A very common mechanism is THERMAL DISSIPATION
What happens is a photon hits an electron, the electron absorbs that energy and reaches an excited state
The electron simply returns to the ground state, and the energy it just absorbed from the photon is released in the
form of HEAT
The energy difference is released in the form of HEAT
Obviously plants dont heat up, so perforin ring systems can effectively avoid this mechanism
Second mechanism is FLUORESCENCE
Excited electron can released that energy in the form of light again
On returning to the ground state that energy is released in the form of light, hv
Most plants dont glow in the dark
Also a mechanism that is not utilized in plants
A third mechanism is ELECTRON TRANSFER
What is electron transfer?
The excited sate electron is in such a high energy state that it is in fact capable of leaving the atom
It moves away, it physically relocates to a different site
The atom that just lost that electron is now oxidized
o It oxidizes into a positive charge
o That electron physically travels to a different location, leaving behind an oxidized atom
The fourth mechanism, and the one that is relevant for this photon harvesting mechanism of photosynthesis is EXCITON
TRANSFER
Exciton transfer is distinct from electron transfer by very simple difference
Electron transfer = the electron physically moves away leaving behind a positively charged particle
Exciton transfer = the energy is directly transferred to a neighboring chromophore, WITHOUT that electron
departing
o That excited state energy is transferred to a second atom, that is right next to it, in the neighborhood
o Upon transfer of the energy, the electron that used to be in the excited state returns to the ground state
o The electron in the ground state of the neighbor goes to the excited state
TRANSFER OF PURE ENERGY, no electrons physically move from one atom to the next, in contrast to
electron transfer
- This exciton transfer is the mechanism by how photons, that have been absorbed by one chromophore, how these photon
travel from one water lily leaf to the next
o Individual chlorophyll molecules cover the surface of the thylakoid membrane
A photon (hv) that hit s a particular chromophore, a perforin ring system, exciting one of the electrons in the
perforin ring system
The energy from the photon is transferred to the neighboring perforin ring system, THROUGH
EXCITON TRANSFER
o And so on
The energy from the sun, hits one of these particles, and it can travel, it can laterally move across
by hopping, by transferring the energy through exciton transfer
o Transferring that energy from one perforin ring system to the next
o AT VERY LITTLE EXPENSE
o Exciton transfer is very efficient
- Whats the purpose of handing around the energy? The hot potato?
o The process is like hot potato
You have 20 people in a room and no one wants to hold the hot potato, so you just hand it around very
quickly
This is pretty much what happens with the photon energy
o The hot potato is handed around UNTIL, this is a random process, sheer chance which perforin ring system picks up
this energy
Handing off of hot potato continues UNTIL sporadically distributed across thylakoid membranes are so
called REACTION CENTERS (energy transduction site)
Exciton energy is handed off from one pigment to the next until it randomly hits one of the reaction
centers
Once it arrives at the reaction center the next stage of photosynthesis takes place
o Think of individual pigments as collection bins

Think of photons hitting leaf surface as rain

All of these pigments would be the funnels that collect the rain and funnels it to the reaction center (Known
as FORSTER/RESONANCE ENERGY TRANSFER)
Once photon reaches the reaction center the next stage can begin
Once these energy components reach the reaction centers we get to stage 2
o In todays plants we have 2 TYPES OF REACTION CENTERS
Referred to as PHOTOSYSTEM II and PHOTOSYSTEM I
These reaction centers are large protein complexes, integral membrane complexes
Photosystem I and II are EMBEDDED in the membrane
In contrast to pigments, they facilitate further chemistry
o The purpose of these 2 photosystems is VERY SIMPLE
Photosystems I and II are responsible for taking the light energy, and converting it into chemical energy
There are 2 light energy inputs in either one of the photosystems
The outputs, there are 2 outputs:
1. The production of NADPH, the redox component
2. The accumulation of PROTONS ON THE LUMEN side of the thylakoid
o What is the lumen? We have the stroma and lumen, what is the lumen?
The disk shaped double membranes, the interior part of the thylakoid
compartments is the lumen
o We generate NADPH but we also accumulate protons inside the thylakoid
These are the 2 mechanisms by which photosystems I and II convert light energy into chemical energy
NADPH is a form of chemical energy storage
These protons that are accumulating can be indirectly used to generate chemical energy in the form
of ATP
o Protons are utilized to synthesize/recycle ATP
If we talk about PI and PII we need to first define a term that is commonly used in connection with photosystem I and II
o That is the REDOX POTENTIAL of a molecule
o What is the definition of REDOX POTENTIAL?
Each chemical entity has a redox potential associate with it, but what do we mean by a redox potential?
What does that tell us?
If I tell you that NADP+ has a redox potential of -0.32 volts what does that number tell you?
The number tells you how likely it is that NADP+ accepts electrons
If you had a molecule of NADP+ and wanted to hand this molecule an extra electron you would
have to FORCE IT because the redox potential is NEGATIVE
It has a tendency of not easily accepting electrons, then what does it do easily?
o IT DONATES electrons easily
If water has a redox potential of +0.82 volts, what does that mean?
Water has a greater tendency to accept electrons, thats why the tendency to accept electrons is
reflected in a POSITIVE redox potential
o Redox potential: the measure of the tendency of a chemical species to acquire electrons and thereby be reduced.
Redox potential is measured in volts (V) or millivolts (mV).
Each species has its own intrinsic reduction potential
The more positive the potential, the greater the species affinity for electrons and tendencies to be reduced
A species with a higher (more positive) redox potential will have a tendency to GAIN electrons from a
species
A solution with a lower (more negative) redox potential will have a tendency to LOSE electrons to a species
o The trick is, in the process of photosynthesis, if you think about the light reaction, what are we actually doing in the
light reaction?
What is the light reaction?
In an abbreviate form: NADP+ + H20 => NADPH + 02
Which of these processes is an oxidation? What happens here exactly?
Youre taking NADP+ and youre dumping electrons into the molecule and making NADPH, what
about the water?
The water actually LOSES electrons, the water gets oxidized, it loses electrons
Looking at the redox potentials, do you think water is happy about giving up electrons based on its redox
potential?
NO! Water has a positive redox potential, it wants to keep/ to accept more electrons

But photosynthesis forces water to give up electrons, is it going to do so freely? NO, it doesnt want
to
At the same time, NADP+ is forced to accept electrons, to form the reduced NADPH
o Does it want to do that? NO!
o NADP+ would rather donate electrons, yet it has to accept those electrons as part of the
chemical storage of the light energy
So, youre looking at an electron transfer reaction that doesnt want to take place
Water doesnt want to let go of the electrons, and NADP+ doesnt want to accept the electrons
How can nature make this process happen nevertheless?
o IT USES LIGHT ENERGY!
It uses the light energy, like a big battery, to force these reactions in photosystem I and photosystem
II
Its like pumping water high up the mountain
Why pump the water so high up the mountain? It is to store that energy, and when we need that
energy we can let the water flow down and spin the turbine to generate the energy
We can use the energy from the sun to pump the water from the high reservoir
The way people represent this, in terms of redox potential and energy storage, describe it as the Z SCHEME
Z-scheme: two-step electron transfer from water oxidation to NADP+ reduction as a function of energy
Water is at bottom, NADP+ is up high
Water is a low energy compound, NADP+ is a high energy compound
Nature, with the help of the photosystems, converts the electrons from water, that are at low energy, converts
those to high energy in the form of NADPH

Why do we need 2 photosystems?

The electrons from water, initially are utilized by PHOTOSYSTEM II
Photosystem II, you can imagine as a pump, driven by sunlight
That pump takes the electrons from water and pumps it up to a higher energy state
If it pumps the electron up to a higher energy state what does that mean?
How does photosystem II actually change the state of those electrons, by moving them to the more
negative redox potential? To the Q state?
What does that mean for that electron?
o It increases the energy of that particular electron
o The electron is now more energetic, it has been taken from water, and with the help of
sunlight has become more energetic
Turns out that this electron, the photosystem II is a very powerful pump, it can inject a lot of energy
into the electron
o But it is not sufficiently powerful enough to pump it up all the way for the energy to be
stored in the form of NADPH
If the first pump is not powerful enough, you can form an intermediate state, and use a second
pump to take it the second step
o In the mountain example: if the first pump does not have enough force you can first form
an intermediate state
o You can use 2 pumps, use a second pump to take it all the way up to the reservoir
Thats why we have to photosystems
PHOTOSYSTEM II: takes the electrons from water, pumps them up, then the electron flow over to
photosystem I

PHOTOSYSTEM I: the second pump, where they get catapulted up to a higher redox potential
o That redox potential is now sufficiently negative, the electron are sufficiently energetic, to
be converted into the NADPH
o To be loaded onto NADP+ to form NADPH
The photosystems are also known as P680 = photosystem II and P700 = photosystem I
P680 and P700 correspond to the energy, the wavelength of light, they absorb

PHOTOSYSTEM II (THE FIRST PUMP)

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What exactly happens in photosystem II? How does it utilize the energy from light and extract the electrons from water? How
does this all happen?
The crystal structure of photosystem II:
o It is an immensely complex and large machinery that constitutes the reaction center
Not just like 3 or 4 alpha helices that are spanning through membrane like bacteriorhodopsin
It is a MASSIVE molecule
o 2 important parts:
1. Stroma: outside of thylakoid
2. Lumen: inside of thylakoid
Have OEC, the oxygen evolution center
What is the chain of events that take place here?
o Lets look at a photon striking a pigment, you have exciton transfer of the energy until it reaches the reaction center,
photosystem II
o You have exciton transfer of neighboring pigment into the reaction center
o The energy that originates from the photon, is now in this photosystem II
Now what happens is instead of the energy being handed from the photosystem II internally you now have
instead of exciton transfer, you have an ELECTRON TRANSFER step
o What happens in photosystem II as energy arrives, the energy is used to take an electron from PII, from the entire
molecule
You take an electron, you pull it out, and you put it on a PLASTOQUINONE (PQ)
Plastoquinone: a small molecule, can absorb electrons very effectively, theyre redox active molecules
As the energy arrives, PII takes that energy and hands off an electron to PQ
o Thereby reducing the plastoquinone
What does that mean to photosystem II, this massive complex? If it physically does an electron transfer to
the plastoquinone?
IT GETS OXIDIZED! It is now positively charged, it just lost an electron
What is its reaction? I WANT MY ELECTRON BACK!
o It has to somehow regain its electron from somewhere
Where can it get electrons from?
o Whats plentiful inside the cell? WATER
o Turns out that this oxidized PII, after it donates that electron to plastoquinone, it turns
around and say I need an electron and it gets that electron from water
Now, how does it do that?
o This is where the OEC, the oxygen evolving center comes in to play

THE OXYGEN EVOLVING CENTER

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In yellow/green/orange are the amino acid molecules of PII

that are used to chelate, to hold in place, a metal oxide
cluster
o Metal oxide cluster consists of calcium, oxygen,
manganese, and tungsten
o For all intents and purposes a manganese-calcium
complex that is at the core of this oxygen evolution center

What this manganese-calcium complex does is it is organized in such a way that it can bind 2 water molecules
o One water molecules binds to the calcium, the other water molecules binds to the neighboring manganese
o 2 water molecules are bound/chelated by this OEC
o Every time a photon strikes and the electron is transferred to a plastoquinone thereby oxidizing PII, it is this OEC
that basically pulls electrons out of these 2 water molecules
o How does it do this? It does this in a stepwise fashion.
Oxygen evolution requires four oxidizing equivalents (photons)
o PS II cycles through 5 oxidation states
o One electron is removed per step
o Final step involves H2O oxidization to O2 + 4H+
Water chelated to Manganese is first fully oxidized to the highly electrophilic OXO complex
o Loses 2 protons
o Every time PII loses an electron, the protein goes
and spits out a proton, and the electron thats left is
kept on the OEC and is reinjected into PII
o PII using OEC, draws electrons out of the water
molecules in order to reduce itself back into its
original state
o When the next photon, the next energy from the
exciton transfer from the pigment arrives, PII can
once again use that energy injected to transfer an
electron to the plastoquinone, thereby being
oxidized again, and then pulling out the next
electron
It takes four photo/light strikes, or 4 exciton transfers, every
exciton transfer pulls out one electron
o 2 water molecules are disintegrated into one
molecules of O2 + 4 protons (H+) and 4 electrons
o Thats the chemistry that takes place
You take 2 water molecules that are bound to manganese and calcium
You pull out 4 protons, one for each light strike
That gives you, as a side product, 4 electrons which are needed to reduce photosystem II
And, by accident you generate molecular oxygen, which is then release by the plant

CYTOCHROM B6F COMPLEX

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What happens to the electrons being dumped?

What happens to the electrons being transferred to the plastoquinone?
o The R group of the plastoquinone is a hydrophobic side chain that allows the plastoquinone to move freely within the
membrane, the hydrophobic environment of the membrane
o When the plastoquinone is fully reduced, it travels from PII to the CYTOCHROME B6F COMPLEX

The cyctochrome B6F complex

o This is a transmembrane protein
o A mediator, redirects flow of electrons
o Uses a bit of the energy from electrons and redirects their flow
o How does it do this?
This protein binds the reduced plastoquinone, which can carry up to 2 electrons
The plastoquinone unloads/releases the 2 electrons once it arrives at the cytochrome b6f complex
It reverses the reaction at PII

Releases 2 electrons and releases to protons to return to its oxidized form

The electrons are transferred into the cytochrome complex
The complex is like a wire, an electrical wire, it allows the electrons to travel through the protein to arrive on the
lumen side
On lumen side is a small protein (PC), plastocyanine
Plastocyanine: a protein that a has a copper center in it that accepts the electrons as they arrive
It picks up the electrons and is released into the lumen
As electrons travel through the cytochrome b6f complex there is also some of the energy is utilized to move
additional protons from stroma
Pumps them into the inside (lumen)
The role of cytochrome b6f complex:
1. It redirects the electron flow
It transfers electrons from PQ onto PC
From small MOLECULE onto the small PROTEIN
2. Also pumps extra protons into the lumen
Plastocyanine:
Relatively tiny protein
At its core is a copper, metal ion chelated
The copper can accept the electron, is reduced, and later on donate that again
Responsible for picking up electron and storing it temporarily

PHOTOSYSTEM I
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PSI transferred excited-state electrons to ferredoxin (Fd)

Fd shuttles electrons to NADP+ reductase for NADPH generation
PII allows for pumping of electrons to intermediate state
o Fully utilization of electrons requires a second pump step: photosystem I
Overall it functions exactly as PII
o Initially the pigment complexes surrounding PI are capturing photons, and through exciton transfer the energy is
transferred to PI
o Knocks out one of the electrons from protein complex
PII: transfers electrons to plastoquinone
PI: transfers electrons to small protein called FERRODOXIN
o Ferrodoxin leaves, leaving behind oxidized form of PI
Where does PI get its electrons back?
o PI USES PLASTOCYANIN which has just accepted an electron from cytochrome b6f complex
o Picks up electron from plastocyanin to reduce itself back into its native state
PI vs. PII
o PII: electron that was kicked off got accepted plastoquinone, electron to re-reduce protein comes from water
o PI: electron gets accepted by ferrodoxin, electron to re-reduce protein comes from plastocyanin
o Otherwise same process, same pumping action
Ferrodoxin is a protein that can now travel to NADP+ reductase, a MEMBRANE-ASSOCIATED enzyme
o NADP+ reductase will bind NADP+ and uses electrons transferred by ferrodoxin to generate NADPH
o Thereby chemically storing the energy that originated from the 2 light strikes