mitochondria guest lecturer: chris moyes, dept of biology contact: [email protected]
TRANSCRIPT
Endosymbiosis
Mitochondria formed as a result of an endosymbiotic event around 2 billion years ago.
From: Gerhart and Kirschner: Cells, Embryos and Evolution
Mitochondrial compartments
Inner membrane•Respiratory chain and ATP synthase•impermeable to most charged molecules•highly folded into invaginations called cristae.
Outer membrane •Permeable to larger molecules
Matrix •Enzymes of the citric acid cycle, mtDNA
Intermembrane space •space between inner and outer membranes
Mitochondrial compartments
Mitochondrial morphology and movement
Mitochondria are dynamic organelles•they may exist as individual organelles•may become elaborate network•move throughout the cell on cytoskeleton
Changes in the network are mediated by fission and fusion proteins
• Dynamin-Related Protein causes fission
• Fuzzy Onion Protein (FZO) causes fusion
Mitochondrial reticulum
Fusion and fission proteins regulate network
Mitochondrial energy production
Three major steps in oxidative phosphorylation
1) Production of reducing equivalents (NADH, FADH2) from glycolysis, fatty acid oxidation, and the citric acid cycle
2) Electron transport and generation of proton motive force
3) Phosphorylation - Synthesis of ATP, driven by the proton motive force
Mitochondria make other products
Mitochondria produce biosynthetic precursors
OXPHOS also leads to the production of:
•Superoxide: formed when O2 steals electrons from the ETC complexes
•Heat: a by-product of the reactions of OXPHOS
Show 14-10, gen overview
Overview of energy production by OXPHOS
Reducing equivalents are produced in the oxidation of carbohydrate
and lipid
Oxidation and Electron Transport
Electrons from NADH and FADH2 are passed down respiratory chain to O2
Electron transport expels protons, creating a proton gradient- the proton motive force (PMF)
Proton motive force (PMF)
The PMF is an electrochemical gradient of membrane potential (ΔΨ) and pH (ΔpH)
The PMF supplies the energy for active transport into the mitochondria
Phosphorylation
The F1Fo ATPase (or ATP synthase) is a molecular motor
-it uses the PMF to make ATP
-it can also be reversed (using ATP hydrolysis to recharge the PMF)
Oxidation and phosphorylation are coupled by a shared dependence on the PMF
Because of this “coupling”, the two processes are interdependent
•If the PMF is large, what would you predict about oxygen consumption?
•If you took away oxygen, what would happen to the PMF?
•What would an increase in [ADP] do to the oxygen consumption?
•What would happen to ATP synthesis and oxygen consumption if the inner membrane became leaky?
Uncoupling proteins
Many mammals warm vital tissues using brown fat
Adipose tissue with abundant mitochondria that possess a the protein thermogenin (or uncoupling protein 1).
UCP-1 short-circuits the proton gradient, increasing VO2 and heat production.
All eukaryotes have proteins related to UCPs, that are thought to prevent the PMF from “over-charging”, thereby reducing ROS production.
Mitochondrial biogenesis requires proteins encoded in 2 genomes (nucleus and mtDNA)
Nucleus
•encode most proteins
•2 copies of each gene per diploid cell
•genes regulated independently
•proteins imported by post-translational import from cytoplasm
mtDNA
•encodes few proteins
•1000’s of copies per cell
•genes transcribed as a polycistron
•transcribed and translated directly in mitochondria)
Peculiarities of mtDNA
mtDNA is a very compact genome-genes attached end to end, with mRNA regions
interspersed among rRNA and tRNA genes-tRNA excision liberates protein-coding genes-many genes lack a full termination codon (TAA)
Diversity-maternal origin (most animals)-many cells have multiple genotypes within a
single cell (heteroplasmy)-defects accumulate with age
Editing of mtDNA polycistron
Nuclear gene expression is coordinated by transcription
factor networks
Mt enzyme synthesis requires coordinated gene expression and
accessory factors