Mitochondrial Atpase
Mitochondrial Atpase - Related Videos
07:38
Measuring In Vitro ATPase Activity for Enzymatic Characterization
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Cited by 27 •
2016
We describe a basic protocol for quantitating in vitro ATPase activity. This protocol can be optimized based on the level of activity and requirements for a given purified ATPase.
02:59
Animal Mitochondrial Genetics
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2020
Among all the organelles in an animal cell, only mitochondria have their own independent genomes. Animal mitochondrial DNA is a double-stranded, closed-circular molecule with around 20,000 base pairs. Mitochondrial DNA is unique in that one of its two strands, the heavy, or H, -strand is guanine rich, whereas the complementary strand is cytosine rich and called the light, or L, -strand. Compared to nuclear DNA, mitochondrial DNA has a very low percentage of non-coding regions and is marked by...
08:40
An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes
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Cited by 38 •
2016
The study of metabolism is becoming increasingly relevant to immunological research. Here, we present an optimized method for measuring glycolysis and mitochondrial respiration in mouse splenocytes, and T and B lymphocytes.
05:53
Mitochondrial Ca2+ Retention Capacity Assay and Ca2+-triggered Mitochondrial Swelling Assay
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Cited by 11 •
2018
This protocol aims to describe a method to examine the Ca2+ retention capacity and Ca2+- triggered mitochondrial swelling of isolated mitochondria of SH-SY5Y cells step-by-step.
09:45
Mitochondrial Isolation from Skeletal Muscle
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Cited by 48 •
2011
This protocol describes a procedure to study the respiration of mitochondria isolated from skeletal muscles. This method was adapted from Scorrano et al. (2007). The mitochondrial isolation procedure requires about 2 hours. The mitochondrial respiration can be completed in about 1 hour.
FAQs
The key coupling event is the conversion of proton flow into rotational motion in the Fo sector. That rotation changes the functional state of the F1 sector, enabling ADP and inorganic phosphate to be combined into ATP. Biological measurements can therefore examine the process at multiple levels, including proton translocation, ATPase activity, or the final ATP-producing outcome.
The respiratory chain establishes the proton gradient that supplies the driving force for the complex. If gradient formation or proton movement changes, rotational coupling and ATP formation can also change. Measuring ATPase activity alongside proton translocation helps connect respiratory-chain function with the performance of the ATP-producing machinery rather than treating ATP levels as an isolated measurement.
ATPase activity reports the catalytic behavior associated with ATP formation, whereas proton-translocation measurements examine movement through the membrane-associated pathway that drives the complex. These readouts address related but distinct stages of the mechanism. Using them together can help determine whether an altered result reflects impaired proton movement, disrupted coupling, or reduced catalytic performance.
Inhibitor sensitivity provides a functional test of how the complex responds to a perturbing compound. Researchers can compare activity or proton translocation under matched conditions with and without the inhibitor, then evaluate the change in response. This approach helps characterize mitochondrial dysfunction or drug effects and can distinguish altered complex behavior from broader changes in energy metabolism.
A study can be organized around one or more measurable outputs: ATPase activity, proton translocation, inhibitor sensitivity, or mitochondrial ATP production. The chosen readout should match the biological question. Activity measurements focus on enzyme performance, proton measurements address membrane-linked energy conversion, and inhibitor comparisons assess how the system responds to a compound or experimental condition.
These measurements are useful when researchers need to evaluate mitochondrial function and energy metabolism under altered conditions. Applications described for the system include studies of metabolic disease, drug effects, cellular responses to mitochondrial dysfunction, and general bioenergetics. Because the assays can examine activity, proton movement, or inhibitor responses, they provide complementary evidence about impaired energy conversion.