Photosynthesis Photosynthesis: process that converts atmospheric CO 2 and H 2 O to carbohydrates Solar energy is captured in chemical form as ATP and NADPH ATP and NADPH are used to convert CO 2 to hexose phosphates Phototrophs: photosynthetic organisms (some bacteria, algae, higher plants) Two Major Reaction Sets Net reaction of photosynthesis is: CO 2 + H 2 O (CH 2 O) + O 2 The oxidation of water is driven by solar energy Electrons from this oxidation pass through an electron-transport chain (which resembles the mitochondrial ETC) Light-dependent Reactions (light reactions) Carbon-assimilation Reactions (dark reactions)

The light reactions Light reactions (light-dependent reactions) H + derived from H 2 O is used in the chemiosmotic synthesis of ATP Hydride ion (H: - ) from H 2 O reduces NADP + to NADPH Release of O 2 from splitting 2H 2 O molecules

The dark reactions Dark reactions (light-independent, carbon assimilation or carbon-fixation reactions) Reduction of gaseous CO 2 to carbohydrate Requires energy of NADPH and ATP Sum of light and dark reactions Both processes can occur simultaneously In the presence of light: H 2 O + ADP + P i + NADPH O 2 + ATP + NADPH + H + Reactions which can occur in the dark: CO 2 + ATP + NADPH + H + (CH 2 O) + ADP + P i + NADP + Sum: CO 2 + H 2 O (CH 2 O) + O 2

The Chloroplast Chloroplasts: specialized organelles in algae and plants where photosynthesis occurs Thylakoid membrane: highly folded continuous membrane network, site of the light-dependent reactions that produce NADPH and ATP Stroma: aqueous matrix of the chloroplast which surrounds the thylakoid membrane Lumen: aqueous space within the thylakoid membrane

Chlorophyll and Other Pigments Capture Light

Recall Energy of a photon h = Planck’s constant = frequency Photons absorbed – electrons go to excited state Can decay back to ground state –Extra energy given off as light, heat, or used to do work (chemical energy) –Direct transfer of energy to a neighboring molecule Exiton Transfer Light-Capturing Pigments Chlorophylls - usually most abundant and most important pigments in light harvesting Contain tetrapyrrole ring (chlorin) similar to heme, but contains Mg 2+ Chlorophylls a (Chl a) and b (Chl b) in plants Bacteriochlorophylls a (BChl a) and b (BChl b) are major pigments in bacteria

Other pigments phycobilins – cyanobacteria & red algae

Accessory pigments – absorb wavelengths that chlorophylls do not Carotenoids – yellow, red or purple Most important one – lutein (yellow) Pigments arranged with specific proteins to form the light-harvesting complexes (LHCs)

Plants usually have twice as much Chl a as Chl b Absorbance spectra complement each other

Photosystems I (PSI) and II (PSII): Functional units of photosynthesis in plants Contain many proteins and pigments embedded in the thylakoid membrane These two electron-transfer complexes operate in series, connected by cytochrome bf complex Electrons are conducted from H 2 O to NADP + PSI and PSII each contain a reaction center (site of the photochemical reaction) Special pair: two chlorophylls in each reaction center that are energized by light (reaction center chlorophylls) In PSI special pair is: P700 (absorb light maximally at 700nm) In PSII the special pair is: P680 (absorb light maximally at 680nm) Light can be captured by antenna molecules (light- harvesting) and transferred among themselves until reaching the special-pair chlorophyll molecules in the reaction center

Diagram of photosynthesis membrane systems Photosystems vary - Photosynthetic bacteria have one-photosystem - Plants & cyanobacteria have 2

Electron Transport in Photosynthesis Distribution of photosynthetic components Overview of Initiation Light capture, electron transport and proton translocation in photosynthesis Light is captured by antenna molecules Light energy drives the transport of electrons from PSII through cytochrome bf complex to PSI and ferridoxin and then to NADPH The proton gradient generated is used to drive ATP production For 2 H 2 O oxidized to 1 O 2, 2 NADP + are reduced to 2 NADPH

Steps to initiate redox chain Absorption of Light Reaction-center chlorophylls contains “special pair” Exiton transfer between antennae molecules Exiton transfer to special pair Energy used to donate an electron Get an electron hole Separation of charge Redox chain can begin

The Z-scheme Z-scheme: path of electron flow and reduction potentials of the components in photosynthesis Absorption of light energy converts P680 and P700 (poor reducing agents) to excited molecules (good reducing agents) Light energy drives the electron flow uphill NADP + is ultimately reduced to NADPH 2H 2 O + 2NADP photons → O 2 + 2NADPH + 2H +

Electron Transport From PSII through Cytochrome bf Electrons for transport are obtained from the oxidation of water (remember the e - hole created) Catalyzed by the oxygen-evolving complex (water- splitting enzyme) of PSII Oxygenic photosynthesis Organisms with only one photosystem do not generate O 2 2H 2 OO H e - Reduction, excitation and oxidation of P680 P680 special-pair pigment of PSII Light excites P680 to P680*, increasing its reducing power e - released to give P680 + P680 + is reduced by e - derived from oxidation of H 2 O

Reduction of plastoquinone to plastoquinol Analogous to Q in the mitochondrial ETC Mobile carrier that goes on to cytochrome bf complex Q-cycle occurs at cyt bf Details: Dimeric protein subunits hold rest of parts together located in granal lamellae (stacked thylakoid membranes) Close to LHCII (light harvesting complex) Electrons pass through only 1 of the dimers e - path: P680 → Pheophytin → PQ A (plastoquinone) → PQ B Net reaction: 4P H + +2 PQ B + 4 photons → 4P PQ B H 2

Splitting of Water Water is split by Oxygen-evolving complex To make 1 O 2 need: –2 H 2 O –Waters release 4 electrons Electrons fill holes in P680 Passed one at a time Mn passes through several oxidation states H + released to the lumen (contributes to pH gradient)

Cytochrome bf links PSI and PSII Similar in function and components to mitochondrial complex III Contains: cytochromes (hemes) and Fe-S protein Has a Q-cycle Is a proton pump – 4H + per PQH 2 Protons pumped: stroma → lumen Proton gradient used to drive ATP synthesis Electrons passed to mobile carrier Plastocyanin

PSI Reduced P700 is excited to P700* (the strongest reducing agent in the chain) by light absorbed by the PSI antenna molecules P700* donates an electron through a series of acceptors to ferredoxin (Fd) e - path: plastocyanin → PSI special pair → A 0 (Chl) → phylloquinonoe (A 1 or Q K ) → several Fe-S centers → NADP + Reduction of NADP + (E o’ = V) by Fd (E o ’= V) is catalyzed by ferredoxin-NADP + oxidoreductase on the stromal membrane side PSI also dimeric – electrons can pass through both Last reaction step 2Fd red + 2H + + NADP +  2Fd ox + NADPH + H +

Arrangement of photosystems PSI stromal lamellae PSII granal lamellae LHCII – holds granal lamellae together varies with phosphorylation of a Tyr adjusts by [PQH 2 ]

ATP Synthesis by Photophosphorylation Photophosphorylation: synthesis of ATP which is dependant upon light energy Chloroplast ATP synthase very similar to mitochondrial counterpart Consists of two major particles: CF o and CF 1 CF o spans the membrane, forms a pore for H + CF 1 protrudes into the stroma and catalyzes ATP synthesis from ADP and P i

About 12 H + passed per O 2 produced –4 from oxygen-evolving complex –Up to 8 from cytochrome bf 4 electrons passed Difference in pH across membrane  pH  3 pH difference is major contributor to free energy stored in the gradient Experimentally shown that ~3 ATP made per O 2 Net equation (non-cyclic photophosphorylation) 2H 2 O + 2NADP photons + ~3ADP + ~3P i → O 2 + 2NADPH + 2H + + ~3 ATP

Cyclic photophosphorylation Produces ATP without also making NADPH Used to adjust ATP/NADPH ratio Uses only PSI Instead of tranferring e - to NADP +, they are passed to plastocyanin Returns e - to reaction center of PSI Still generate pH gradient and ATP Net reaction: ADP + P i + light  ATP + H 2 O Summary of ATP Synthases