Mitochondria and chloroplasts Flashcards
Proton electrochemical gradient
The proton electrochemical gradient across a membrane is generated by energy released as electrons travel down their electric potential gradient through the electron transport chain. This energy is eventually used to synthesize ATP
Proton-motive force
The energy stored in the proton electrochemical gradient, used to power ATP synthesis
ATP synthase
The ATP synthesizing enzyme. Used to move protons down their electrochemical gradient. As the protons move down their gradient, they release energy used to make ATP from ADP and Pi
Aerobic respiration stage 1
Stage 1 is glycolysis, which occurs in the cytoplasm. One glucose molecule is used to produce 2 pyruvate molecules. 2 ATPs and some NADH are also produced
Aerobic respiration stage 2
Stage 2 is the citric acid cycle, which occurs in the mitochondria. Pyruvate is oxidized to produce the necessary starting material (acetyl CoA). Produces carbon dioxide, ATP, and electron carriers NADH and FADH2, which store energy for later
Aerobic respiration stage 3
Stage 3 is the electron transport chain. Electron carriers (NADH and FADH2 transfer their electrons to molecules near the beginning of the transport chain. As electrons are passed down the chain, they move from a higher to a lower energy level, releasing energy. Some of the energy is used to pump H+ ions, moving them out of the matrix and into the intermembrane space. This pumping establishes an electrochemical gradient. At the end of the electron transport chain, electrons are transferred to molecular oxygen, which splits in half and takes up H+ to form water.
Aerobic respiration stage 4
Stage 4 is ATP synthesis. As H+ ions flow down their gradient and back into the matrix, they pass through an enzyme called ATP synthase, which harnesses the flow of protons to synthesize ATP.
Stage 1 of photosynthesis
Occurs in the chloroplast- energy absorption by pigments and direct transfer to electrons. Chlorophyll absorbs the energy, entering an excited state. It donates an electron to the adjacent chlorophyll, which passes it down a chain of acceptor molecules. Quinone is the primary electron acceptor, similar to coenzyme Q in the mitochondria.
Stage 2 of photosynthesis
Electron transport and proton motive force (H+ gradient). Electrons move from the high energy electron acceptor QH2 to a proton pump (the cytochrome bf complex).
Plastocyanin
Plastocyanin is an electron carrier protein that transfers electrons from the cytochrome bf complex to a complex called photosystem I. The electrons are low energy after going through the proton pump.
Stage 3 of photosynthesis
The proton gradient across the thylakoid membrane, which was generated by light, is used to synthesize ATP. Protons move down their concentration gradient from the thylakoid lumen to the stroma through the chloroplast F0F1 complex (ATP synthase).
Stage 4 of photosynthesis
Carbon fixation. The NADH and ATP produced in stages 2 and 3 drive the synthesis of six-carbon sugars from water and carbon dioxide. Starch is synthesized in the stroma.
Energy source for aerobic respiration
The chemical bonds of fuel molecules- sugars (primarily glucose) and fatty acids.
Energy source for photosynthesis
Photons (sunlight)
What is unique about Henneguya
salminicola?
It can survive without oxygen, which is changing the definition of what an animal can be. The 10 celled Henneguya
salminicola lives as a
parasite in salmon and is
related to jellyfish. It has lost its mitochondrial genome. Not sure how it survives but it probably steals nutrients from host.
Super resolution microscopy of the mitochondria
The stacks of the cristae and their membranes can be visualized this way. The cristae are invaginations of the inner mitochondrial membrane and act to expand the surface area of that membrane. This is where ATP synthesis occurs.
Microtubules and the mitochondria
Microtubules are essential for mitochondrial movement, as well as fission and fusion. While actin may not be a major highway for mitochondrial movement it may function as an entrance ramp, helping mitochondria get to the highway as needed. Microtubules can then move the selected mitochondria and regulate fission and fusion events. This model is similar to mitochondrial behavior in neurons. Neurons utilize the actin cytoskeleton to move mitochondria shorter distances and microtubules for long distance transport
Internal structure of the mitochondria
There is a smooth outer membrane that forms the outside boundary of the mitochondria. The inner membrane is continuous and composed of 3 domains. The cristae are invaginations from the inner membrane to the center of the mitochondria. The intermembrane space is the fluid filled space between the inner and outer membranes. The matrix is the fluid filled space within the inner membrane, and it contains mtDNA, ribosomes, and granules.
Transmission electron microscopy of the mitochondria
The mitochondrial membranes can be observed using this imaging technique. Crista junctions (that separate the crista membranes from the inner boundary membrane) can be seen.
Mitochondrial microprotein
New “microprotein” (54 AA) called PIGBOS discovered on the outer mitochondrial membrane (green fluorescence). It communicates with the ER and regulates the UPR via
the ER protein CLCC1 at mitochondrial ER contact sites. Loss of PIGBOS increases UPR leading to
increased cell death
Unfolded-protein response
Cells respond to the presence of unfolded proteins in the rough ER by increasing transcription of genes that encode ER chaperones and folding catalysts.
Mitochondria associated membranes (MAMs)
Regions of direct contact between the ER membrane and the outer mitochondrial membrane. They affect mitochondrial structure and function. For example, in response to elevated IP3, calcium moves through the MAM into the mitochondrial matrix. Elevated calcium levels in the matrix increase ATP synthesis.
The inner mitochondrial membrane (IMM)
The IMM contains the ETC and ATP synthase and an unusual four legged phospholipid called cardiolipin (CL)
which is now widely studied.
Cardiolipin
Cardiolipin interacts with and is required for optimal activity of several IMM proteins, including the enzyme complexes of the electron transport chain (ETC) and ATP production and for their organization into supercomplexes. Acts like “glue” for the ETC. CL also plays an important role in mitochondrial membrane morphology, stability and dynamics, in mitochondrial biogenesis and protein import, in mitophagy, and in different mitochondrial steps of the apoptotic process
Fusion and fission
The mitochondria undergo frequent fusions (merging) and fissions (breaking apart), which can generate tubular or branched networks. This could account for the variety in mitochondrial morphologies seen across cell types
How is mitochondrial fusion regulated
Integral membrane proteins MFN1 and MFN2 mediate outer mitochondrial membrane fusion. This is followed by fusion of the inner mitochondrial membranes, which is mediated by integral membrane protein OPA1.
How is fission regulated?
A GTPase called DRP1. Post-translational modifications of DRP1 regulate fission
Tunneling nanotubes
Tunneling nanotubes are projections of the cell membrane that connect the cytoplasm of animal cells. They act like gap junctions by transferring chemical and electrical signals between the cells.
In addition to intracellular movement, mitochondria and mtDNA have been found to be transferred from one cell to another via these membrane tubules
Mitochondrial permeability transition pore
A transmembrane protein residing in the mitochondrial inner membrane. Normally closed, this large protein pore opens when stimulated by mitochondrial matrix Ca2+ accumulation, adenine nucleotide depletion, increased phosphate concentration or oxidative stress. Opening of the pore is linked to apoptosis.
Why does mitochondrial fission occur? (2)
- Distribute mitochondria into 2 daughter cells
- Eliminate damaged parts of the mitochondria
Intrinsic triggered apoptosis
Mitochondria can trigger cell death in response to intrinsic cell stress- increase in reactive oxygen species. Chemotherapy and radiation creates mutations in the nucleus so the cell is triggered to go into apoptosis
What happens when the MPTP is opened? (4)
Triggers the mitochondrial apoptotic cascade
1. Cytochrome C is located in the ETC- triggers apoptosis when released
2. Next, cytochrome C is in the cytoplasm, binds to APAF1
3. APAF-1 now binds to procaspase 9- caspase 9 is inactive
4. Caspase 9 binds to procaspase 3- as a result we now have caspase 3 (executioner enzyme that kills the cell)
Extrinsic apoptotic cascade
Linked via “cell death receptors”- activates caspase 8- then activates caspase 3.
Bcl-2
Anti apoptotic protein (prevents the mitochondria from triggering apoptosis)- named because of discovery in B cell lymphoma. Overproduction of Bcl-2 is very common in many different cancers and can make them resistant to chemotherapy and radiation
Treating Bcl-2 overexpressing cancers (2)
2 new drugs are being tested.
1. Genasense – Bcl2 antisense oligonucleotide
targets Bcl2 mRNA
2. Obatoclax – Small molecule Bcl-2 inhibitor
Venetoclax
Can now be used for a broader group of patients with chronic lymphocytic leukemia (CLL). Venetoclax was initially approved by the FDA in 2016 to treat people with CLL that has a specific genomic alteration, called deletion 17p. It targets BCL‑2 proteins and attaches to them. When Venetoclax is attached to these proteins, it helps to restore the process of apoptosis, allowing these cells to self‑destruct.
Synta Pharmaceuticals
In preclinical trials with
Elesclomol – a drug that triggers apoptosis through
targeting the electron transport chain in cancer cell mitochondria. Fast Track Orphan Drug Status for metastatic melanoma. Mechanism- increases oxidative stress
Reactive oxygen species
Highly reactive oxygen containing molecules. They are by products of the ETC and can be purposefully generated by WBCs to kill pathogens. ROS can react with and damage important biological molecules, such as lipids, proteins, and DNA