Mitochondria (1) Flashcards
Which transporter exchanges ATP for ADP, moving ATP to the cytosol, and where is it located?
Ant transporter
Inner membrane
What does an O2 electrode do?
Detects rate of O2 removal from surrounding buffer (must be airtight) - measures O2 consumption
What does a CCCP lipophilic proton shuttle do?
Sits in mitochondrial membrane - removes H+ gradient across membrane
Complexes I-IV pump down electrochemical gradient
Measure maximum respiration rate of mitochondrion
What does a seahorse assay do?
Measures O2 uptake of cells
Add drugs to see effect - e.g. oligomycin
What does oligomycin do?
Inhibits ATP synthase - H+ gradient too steep for complexes I-IV - slows respiration
What is TMRM and what does it do?
Lipophilic cation
Measures mitochondrial potential - accumulates in cell compartments with negative charge - taken up by mitochondria
What is the use of NAD(P)H/FAD2+ fluorescence?
Illuminate tissue with light - fluorescence intensity corresponds to amount of NADH/NADPH/FAD2+
Changes in fluorescence show changes in redox state - shows O2 consumption and substrate supply
What type of light is used to visualise NAD and what is the resulting fluorescence?
UV light
NADH - blue fluorescence
NAD+ - no fluorescence
What type of light is used to visualise FAD and what is the resulting fluorescence?
Blue light
FAD2+ - green fluorescence
FADH2 - no fluorescence
What are the mtDNA copies in each mitochondrion packaged as?
Nucleoids
What is mitochondrial heteroplasmy?
Some mitochondria in cell have wildtype mtDNA - some have mutant mtDNA - role in varying mitochondrial disease severity
What is biogenesis?
Mitochondrial replication
What is the process of mitophagy?
Autophagosome membrane wraps around depolarised mitochondrion
Autophagosome fuses with lysosome
Mitochondrion degraded by hydrolysis
Which protein regulates the fusion of the inner mitochondrial membrane?
OPA1
Which protein regulates the fusion of the outer mitochondrial membrane?
Mfn1/2
