- #1
JamesPhD
- 17
- 3
I purposely added this post to the general physics thread rather than the biology thread because the solution is probably not in biology.
Basically my PhD is in mitochondrial biology. Mitochondria are the energy producing organelles in the body which make energy by adding a chemical bond converting a di-phosphate ADP to ATP (tri-phosphate).
The problem is the precise machinery about how this works is not entirely known. The oxidative phosphorylation system which is composed of 5 large protein complexes known as complexes I-V are responsible for making ATP which is shown in any high school textbook. These mitochondria have double membranes and can be isolated from cells and tissues and treated like individual cells on their own.
However, in the past 20 years, through the use of less volatile detergents used for breaking the mitochondrial membrane, we have learned that these individual complexes can combine together in a dynamic way under different metabolic cues to fine tune the system and make mitochondria more efficient. This happens in exercise for instance.
But the act of breaking the membrane may shatter this protein machine into different 'parts' and the parts are what we see. These parts are called supercomplexes.
So how could we see these protein-protein interactions intact? People used formaldyhyde to cross link proteins and using this method found what they called the megacomplex, larger than supercomplexes.
But I think that these complexes all work together in a respiratory string, which makes ATP but we just can't see it.
I was considering using antibodies to view these complexes however antibodies are 1500 Da while proteins 100 times smaller in size are the ones which penetrate the membrane. So the act of using antibodies would breach the membrane.
Maybe the use of a hypotonic solution in which mitochondria uptake into their core might work. Then we could spin the isolated mitochondria down, transfer them into another solution and trigger the solidification of the uptaken hypotonic solution which would leave an imprint of proteins on the inner mitochondrial membrane and viewed using a microscope such as an electron-microsope?
It would be one of the most important questions to answer in biology, how does this machine work. But I don't think the answer is in biology. Perhaps there is some tool in physics or engineering that people routinely used and could be applied to this question. I need a non-biologist perspective. I appreciate your input.
Basically my PhD is in mitochondrial biology. Mitochondria are the energy producing organelles in the body which make energy by adding a chemical bond converting a di-phosphate ADP to ATP (tri-phosphate).
The problem is the precise machinery about how this works is not entirely known. The oxidative phosphorylation system which is composed of 5 large protein complexes known as complexes I-V are responsible for making ATP which is shown in any high school textbook. These mitochondria have double membranes and can be isolated from cells and tissues and treated like individual cells on their own.
However, in the past 20 years, through the use of less volatile detergents used for breaking the mitochondrial membrane, we have learned that these individual complexes can combine together in a dynamic way under different metabolic cues to fine tune the system and make mitochondria more efficient. This happens in exercise for instance.
But the act of breaking the membrane may shatter this protein machine into different 'parts' and the parts are what we see. These parts are called supercomplexes.
So how could we see these protein-protein interactions intact? People used formaldyhyde to cross link proteins and using this method found what they called the megacomplex, larger than supercomplexes.
But I think that these complexes all work together in a respiratory string, which makes ATP but we just can't see it.
I was considering using antibodies to view these complexes however antibodies are 1500 Da while proteins 100 times smaller in size are the ones which penetrate the membrane. So the act of using antibodies would breach the membrane.
Maybe the use of a hypotonic solution in which mitochondria uptake into their core might work. Then we could spin the isolated mitochondria down, transfer them into another solution and trigger the solidification of the uptaken hypotonic solution which would leave an imprint of proteins on the inner mitochondrial membrane and viewed using a microscope such as an electron-microsope?
It would be one of the most important questions to answer in biology, how does this machine work. But I don't think the answer is in biology. Perhaps there is some tool in physics or engineering that people routinely used and could be applied to this question. I need a non-biologist perspective. I appreciate your input.
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