10: Chloroplast Structure and Function
an excited state
Heterotrophs are those than depend on an
external source while autotrophs can
manufacture their own nutrients
Chemoautotrophs utilize chemical energy in
inorganic molecules
Photoautotrophs utilize the energy from the
sun
During photosynthesis, low-energy electrons are
removed from a donor compound and converted
into high-energy electrons using the energy
absorbed from light
The switch from sulfur to oxygen was hard since
sulfur has lower affinity for electrons
Polypeptides found in modern-day chloroplasts
are encoded by both the chloroplast and nuclear
genome
10.1 Chloroplast Structure and Function
Located predominantly in the mesophyll cells of
leaves
Lens-shaped in higher plants, approx. 2-4 x 5-10
m, 20 per cell
Arise from fission from preexisting chloroplasts or
from non-pigmented precursors called
proplastids
Were identified as the site of photosynthesis in
1887 by T. Engelmann upon observing that
bacteria would aggregate near the chloroplast
Contains two membranes separated by a narrow
space
o Outer membrane contain porins
o Inner membrane is highly impermeable;
substances need the help of transporters
An internal membrane system separate from the
bi-layer is organized into sacs called thylakoid,
stacked into grana
o Space inside the thylakoid is the lumen
o Space within the chloroplast is the stroma
Stroma contains small double-stranded circular
DNA and prokaryotic-like ribosomes
Thylakoid membranes have high protein content
and low phospholipid but instead have high
percentage of galactose-containing glycolypids
10.2 An Overview of Photosynthetic Metabolism
Photosynthesis essentially is a redox reaction
o H2A (S/O) is an electron donor; reducing
agent
o CO2 is an oxidizing agent
Each molecule of O2 is derived from the
breakdown of two molecules of H2O
Photosynthesis is then the reverse of
mitochondrial respiration
o Mitochondrial respiration reduces oxygen to
water
o Photosynthesis oxidizes water to oxygen
light-dependent reactions: energy from the sun is
absorbed and stored as ATP and NADPH
light-independent reactions: carbohydrates are
synthesized using the stored energy
10.3 The Absorption of Light
light travels in energy called photons; the energy
depends on the wavelength of light
o The shorted the wavelength, the higher
energy content
When a photon is absorbed by a molecule, an
electron becomes too energetic to be pushed
from an inner to a higher/outer orbital shifting to
In isolated chlorophyll, the solution becomes
fluorescent because the absorbed energy is re-
emitted at longer (low energy) wavelength
In isolated chloroplasts, only a faint fluorescence
is observed. Very little of the energy absorbed is
dissipated but are instead transferred to electron
acceptors
Photosynthetic Pigments
Pigments appear colored because they only
absorb a light at a specific wavelength
Chlorophyll absorbs strongly in the blue and red
Chlorophyll structure:
o Porphyrin ring with Magnesium for light
absorption
Conjugated (alternating single and
double bonds)
Strong absorber of light
Absorbed energy causes a redistribution
of the electron density of the molecule
o Hydrophobic phytol chain that keeps the
chlorophyll embedded in the membrane
Carotenoids absorb well in the blue and green
spectrum; act as secondary light collectors
during photosynthesis and draws excess energy
from chlorophyll as heat
10.4 Photosynthesis Units and Reaction Centers
Not all chlorophyll molecules in a chloroplast are
involved in conversion of light into chemical
energy
Chlorophyll molecules act together as one
photosynthetic unit in which only one member of
the group, reaction-center chlorophyll
actually transfers electrons to an electron
acceptor
These pigment molecules act as an antenna that
absorbs photons of various wavelengths and
transfers that energy (excitation energy) to the
reaction-center
Energy can only be transferred to an equal or
less energy-requiring molecule
Oxygen Formation: PSI & PSII
Light-absorbing reactions take place in protein-
complexes called photosystems
Photosystem II (PSII) boosts the electron to a
midway point; P680
Photosystem I (PSI) raises the electrons from a
midway point to an energy level above NADP+
After losing electrons, PS becomes positively
charged which attracts electrons
Z Electron flow: water PSII PSI NADP+
Electron transfer releases energy which is used to
establish a proton gradient to drive the synthesis
of ATP needed for the synthesis of carbohydrates
PSII Operations
PSII uses absorbed energy to remove electrons
from water and generate a proton gradient
D1 & D2 bind the P680 reaction-center with other
cofactors
Light-harvesting complex II (LHCII) contain the
antenna and are situated outside the core of the
photosystem; can migrate along the thylakoid
membrane
Formation of a single oxygen requires 4 photons
to be absorbed by PSII

PSII Platoquinone
1. P680 transfers received electron to pheophytin,
the primary electron acceptor creating a
separation of charge
P680+: oxidizing agent
Pheo-: reducing agent
2. Pheo- transfers its electron to plastoquinone
bound near the membrane
3. Electron from PQA is then transferred to another
PQB to produce a semi-reduced molecule that
remains firmly bound to the D1
4. P680 is reduced back
5. 2-3 is repeated forming plastoquinol from the
joined molecules (PQH2). The protons used are
derived from the stroma (going in) and thus
creates a gradient
6. PQH2 then diffuses into the lipid bilayer
Water PSII
The redox potential of P680 is strong enough for
photolysis of water
Formation of one molecule of oxygen requires 4
electrons from 2 molecules of water
Oxygen-evolving complex: 4Mn + 1Ca
accumulates four oxidizing equivalents by
transferring 4 electrons, one at a time to PSII
From PSII to PSI
1. PQH2 is mobile and binds to cytochrome b6f
which engages in the Q cycle
2. PQH2 donates 2 electrons to cytochrome b6f and
translocate 2 protons to the lumen
3. cytochrome b6f translocate a net of 4 protons
4. Electrons are transferred to plastocyanin
situated on the luminal side which carries
electrons to PSI
PSI Operations
PSI is made of a core + LHCI
1. Energy passed to reaction center
2. Electrons are passed to chlorophyll a, the
primary electron acceptor creating a separation
of charges: P700- and Ao- (reducing agent)
3. Ao is passed to phylloquinone and then to 3
iron-sulfur clusters
4. Electron is transferred out via ferredoxin
NADP+ reduction is done via ferredoxin-NADP+
reductase which requires 2 ferredoxin molecules
Not all electron passed to ferredoxin ends up in NADP
reduction. Some are used for other reductions.
Photosynthetic Electron Transport
Electrons travel from water to NADP+ via the 2
photosystems
PSII produce Oxygen
PSI produce NADP
Each photosystem requires 4 electrons; net of 8
electrons are needed
Proton gradient is formed via:
o Splitting of water
o Oxidation of plastoquinol
o Reduction of NADP+ and PQ
Practical Application: Killing Weeds
Bind to open Q site, blocking transport through
PSII
Paraquat competes for electrons with ferredoxin
but also creates free radicals
10.5 Photophosphorylation
Conversion of one mole of CO 2 to one mole of
carbohydrate requires 3 moles of ATP and 2
NADPH
ATP synthase
o Head: CF1 which projects outward to the
stroma
o Base: CF0
Protons move from the lumen to the stroma
Movement of protons during electron transport is
neutralized by the movement of other ions. Thus,
proton gradient is mainly due to pH only
Non-cyclic phosphorylation: electrons move
linearly from water to NADP+
Cyclic phosphorylation: carried out by PSI
independent of PSII
1. PSI absorbs light passed to ferredoxin
2. Electron is routed back to the electron-
deficient reaction-center
**cyclic phosphorylation drives enough energy to
build the proton gradient necessary**
10.6 Carbon Dioxide Synthesis
Melvin Calvin: radioactive Carbon; killed cells
immediately (stopping enzyme activity) and
analyzed products via chromatography
C3 plants/Calvin Cycle
First molecule to be discovered was 3-
phosphoglycerate (PGA)
RuBP + CO2 6 Carbon fragmented into 2
molecules of 3-PGA
Catalyzed by ribulose bisphosphate carboxylase
oxygenase (rubisco) which is only capable of
fixing three molecules of CO2 per second. This is
the lowest turnover for any enzyme so plants
contain as much as half of its proteins as Rubisco
making it the most abundant protein on earth
Cycle comprises of 3 main steps
o Carboxylation of RuBP forming PGA
o Reduction of PGA to sugar by formation of
glyceraldehyde 3-phosphate (GAP) using
NADPH and ATP
o Regeneration of RuBP via ATP
For every 6 molecules of CO 2 , 12 GAP are
produced
o 10 are recycled to form 6 RuBP
o 2 are products
GAP can be:
o Exported to the cytosol in exchange of
phosphate ions to synthesize sucrose
(~glucose)
o Remain in the chloroplast where it is
converted to starch (~glycogen)
Redox Control
Light dependent
Controlled by thioredoxin which reduces
disulfide bridges into sulfhydryl groups in
enzymes causing their activation
Some of the electrons go to thioredoxin instead
of ferredoxin
During the dark, thioredoxin is no longer reduced
by ferredoxin and the enymes revert back to an
oxidized state
Photorespiration
Glycolate is a product of a reaction catalyzed by
Rubisco where Oxygen attaches to RuBP forming
2-phosphoglycolate which is converted to
glycolate
Glycolate is transferred to the peroxisome where
CO2 is released
Upatake of oxygen and release of carbon dioxide
Waste of plants energy
Occurs when the concentration of oxygen is high
as compared to carbon dioxide (when plants
close stomata to prevent water loss)
Rubisco only has modest preference for carbon
dioxide vs oxygen making this very easy since
neither of the two binds to the active site of the
enzyme
C4 plants
CO2 + phosphoenolpyruvate (PEP) via
phosphoenolpyruvate carboxylase
Tropical grasses that are adapted to hot and arid
environments that can close stomata to conserve
water but can still maintain photosynthesis;
wont work well in cooler temperatures or in
higher altitudes
Continues net synthesis of carbohydrate until CO 2
levels have dropped to 1-2 ppm
PEP carboxylase can operate at low levels of CO2
and is not inhibited by oxygen
Downside is it requires more ATP and NADPH
Happens in different parts of a leaf
o Mesophyll cells where fixation occurs
o Bundle-sheath cells sealed off from
atmospheric gass
Evolved separately from C3 ancestors
CAM plants
Also utilize PEP carboxylase but carry out light-
dependent reactions in different times of the day
Keep stomata closed at day, opened at night
Malate is evident as sour morning taste