Proline Inhibits Mitochondria

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
Some weeks ago, I had what I think was a viral infection, and it made my ME symptoms worse after high-protein meals. After the infection was gone, I remained sensitive to proteins. After much experimentation, I determined that I was sensitive to proline. If I consume ~1400 mg of proline, I'll get a noticeable increase in ME symptoms, most noticeably brainfog and muscle aches (I assume neuropathic). Larger doses of proline makes the severity much worse. The time for the decrease in symptoms depends on the dose: just past the threshold may take hours; much larger doses can take days. My response to proline seems a lot like PEM, which in turn seems a lot like an increase in my usual ME symptoms, so this might provide an insight into the core dysfunction of ME and/ or PEM.

Why might proline be involved in ME? Well, proline is critical for mitochondrial function. This paper explains it: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4752965/

"Proline dehydrogenase (PRODH), a.k.a. proline oxidase (POX), is a mitochondrial inner membrane protein. Oxidation of proline to pyrroline-5-carboxylate (P5C) mediates the proline cycle that shuttles NADP(H)/NADP(+) redox equivalents between mitochondria and cytosol and forms a metabolic interlock with the pentose phosphate pathway"

"Proline dehydrogenase/oxidase (PRODH/POX) is a mitochondrial protein critical to multiple stress pathways."

"We found that PRODH/POX binds directly to CoQ1 and that CoQ1-dependent PRODH/POX activity required functional Complex III and Complex IV. PRODH/POX supported respiration in living cells during nutrient stress; however, expression of PRODH/POX resulted in an overall decrease in respiratory fitness. Effects on respiratory fitness were inhibited by DHP and NAC, indicating that these effects were mediated by PRODH/POX-dependent reactive oxygen species (ROS) generation. PRODH/POX expression resulted in a dose-dependent down-regulation of Complexes I–IV of the ETC, and this effect was also mitigated by the addition of DHP and NAC."

To me, it certainly seems like it's possible for proline to be involved in the cellular dysfunction found in ME. I came across a couple of papers that showed that excess proline can reduce mitochondrial function in the brain, an effect that can be reduced by pre-treatment with antioxidants. I'll have to experiment with that to see if it reduces my sensitivity.

Abnormal proline metabolism might explain the collagen problems some PWME report, since proline is critical for collagen formation. Alternatively, maybe a problem with maintaining collagen worsens ME by releasing more proline to muck up mitochondria.

There's a human brain-specific high affinity L-proline transporter (hPROT). "These findings warrant the consideration of a synaptic regulatory role for this transporter and its presumed natural substrate, L-proline, in the mammalian central nervous system." That sounds like it could be one of those biological mechanisms that no one notices when doing ordinary medical testing, but whose subtle dysfunction could result in significant symptoms.

I assume that my present sensitivity to proline is unusual, so I don't think that ME is proline sensitivity. However, this link between proline and ME/PEM symptoms might be worth investigating further, so I'm posting this to see if it triggers some ideas in other people, or makes some researcher evaluate test results from a new perspective.

Also, for people who are wondering why some foods make them feel worse, you could check to see if they're proline-rich. Wheat is very high in protein (there goes another staple from my diet :(). Peanuts are too, and of course, meats. Proline sensitivity is very annoying, diet-wise. :grumpy:


One suggestion for a simple test for researchers: do the nanoneedle test with extra proline added to the fluid. Doing it with extra antioxidants and extra proline might be worthwhile too.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
Just a n=1 anecdote:
Yah, I don't think ME involves elevated serum proline, since plenty of people do better on a high-protein diet. It could, however, be a problem with the metabolism inside cells, or transport across the membrane, or some other such thing. My case might be unique, involving some additional dysfunction that makes me sensitive to serum proline level. The fact that it's inducing ME/PEM-like symptoms makes me suspicious of it. Of course, it's possible that my proline issue is not directly related to ME, but affects some other subsystem which in turn affects my ME.

It sure would be nice if someone out there said: "Hey, I think I know why it's doing that." :)
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
On Monday, I had to go into town for a visit to the dentist (crown prep). That seems like the kind of trip that should give me PEM. The road was even slippery with mud, for extra stress. Thus I was expecting PEM.

None occurred. Maybe my present low-proline diet made the difference? I don't have any more reliable methods of inducing PEM, so it's hard to test that hypothesis further, but I thought I'd mention it. Maybe diet affects other people's PEM severity or triggering threshold.

The low-proline diet is also still letting me sleep longer between wakings. I've had several sleeps up to 5.5 hrs long, which is amazing after years of waking every 90 minutes. I don't feel more refreshed, but it's still nice to not wake that much, and it's probably healthier.

Luckily, proline isn't an essential amino acid, so I probably don't have to worry about deficiency. The diet cuts out a lot of enjoyable foods though. Such is life with ME. :(
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
What does your diet look like without prolein?
Cornstarch pancakes for two meals, for calories without problematic nutrients. Afternoon meal might be quinoa with onions and carrots. Some potato, rice or wheat (limited) snacks, plus milk in coffee or hot chocolate. Yes, very boring, and I wonder if it will drive me insane at some point, and I'll go chase down a bunny and eat it. :wide-eyed:

For protein, I'll have to figure out something for long-term, but I'm still in a 'seeing what happens' phase, and I'm not expecting any signs of protein deficiency any time soon. My legs feel a bit wobbly since the diet chance, so I think I'm missing something that I need, so it's more experimentation for me.

Also you say this proline sensitivity appeared after a recent infection yet wasnt here before?
I think I may have had a slight sensitivity to it for a year or more before that, but I'm not certain. I had been wondering whether meat was worsening my symptoms, but I didn't do a definite controlled experiment. My reaction to protein during and after the 'viral infection (if that's what it was)' was dramatically more severe.

My frequent wakings started up several years ago, so if that was due to proline sensitivity, I've had part of the problem for a long time.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
I noticed that soup, made with bouillon cubes, was also triggering more brainfog, even though the ingredients didn't seem that high in proline. It does have MSG though, and I found out that our bodies convert glutamine into proline, so that probably explains it.

There is a rare genetic disease of elevated proline. It's usually asymptomatic, so that's probably not what I have. I could go through the hassle of getting my serum proline measured, but that doesn't lead to any useful actions, since there's no treatment for excess proline. If this is an unusual sensitivity to proline, then it likely involved whatever chemical reactions are involved in ME, which means that no one would know what the reaction was or what to do about it.

I'll see what the local heath store has in the way of essential amino acids without proline (or glutamine), or else order some online. Maybe boosting those other amino acids will help.
 

sb4

Senior Member
Messages
1,599
Likes
2,829
Location
United Kingdom
@Wishful An idea for an experiment would be to get amino acids without protein and see if you notice anything, then get proline on its own and take a decent dose to see if negative symptoms appear.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
The symptoms correlate very strongly with proline content of the foods, so I don't see it worthwhile buying pure evil (proline) just to feel extra lousy. I will try the essential amino acids to see whether maybe they'll affect my inner amino acid balancing mechanisms. The production of the various enzymes and whatnot do vary with diet.

If the essentials have an effect, I can also try the other non-essentials.

This dramatic change in proline sensitivity happened after what seemed to be a viral infection. Maybe the virus left behind some extra baggage that is messing with my proline.
 

vision blue

What big eyes...
Messages
1,643
Likes
2,128
It sure would be nice if someone out there said: "Hey, I think I know why it's doing that." :)
Ever been tested for antibodies to alpha-gal? that can give you weird responses to certain proteins, especially red meat but its more wide spread than that. that would be most especially true if you got it after an infection since thats when alpha gal antibodies seem to start.

How were you able to narrow it down to proline? for example, you said you react to bouillin and as you mentioned that also has high glutamate. Did you test a couple dozen things and only common demoniator was proline? like which things?

i'm a fan of amino acid testing, ideally both blood and urine the same day (I've done this!). besides seeing how proline is doing on both of these (they trade off), you can also see if the culpit is really upstram or downstream of proline.

not sure its effects on mitochondira impress me. for any amino acid you pick, you can probably find a fistful of papers tying it to some effect on the mitochondria.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
Ever been tested for antibodies to alpha-gal?
No, my responses don't fit that.

Did you test a couple dozen things and only common demoniator was proline?
That's more or less correct. Not a couple dozen, just under a dozen. It was actually quite hard to find foods that varied significantly in amino acid ratios. The ones with a higher ratio of non-proline/proline rules those out as culprits. Then I chose two slices of bacon as my baseline for triggering symptoms, and tried several other foods at amounts that contained the same amount of proline. Those all gave the same level of symptoms at that amount, and no symptoms at significantly lower amounts. It would be nice if I found a food with a significantly lower ratio of proline to the other aminos, but the best I've found is garlic, and even that isn't all that much lower in proline.

i'm a fan of amino acid testing,
I don't see any potential for useful results. If it's highly elevated, I can't do anything about that. If it's not elevated, I can't do anything about that either.

I'm not convinced that it's effect is via mitochondria either. Apparently proline is also neurotoxic. It's also involved in all sorts of other cellular functions, and viral replication. Just another extension of the mystery of ME.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
On my trip to town yesterday, I decided that I was desperate for some meat, so I bought some fried chicken legs. My tummy was much happier with that. :thumbsup: An hour and a half after eating it, I started feeling a bit dizzy, which got bad enough that I had to lie down. The worst passed in a couple of hours. This morning I'm still a bit brainfogged, but managed a good walk and probably feel no more brainfogged than without the meat. So, this isn't quite as simple as symptoms being directly proportional to proline. That leaves room for hope. Maybe another amino acid or fatty acid helps counteract the proline. Maybe gut function or microbiome is involved. I don't mind having to experiment with meat-filled meals. :)

I hope no one finds out that their 'thing to experiment with' is some food they really hate. :yuck:
 

vision blue

What big eyes...
Messages
1,643
Likes
2,128
So far the specifics you mentioned are bouillion cubes, bacon, and froed chicken legs. All 3 of these also have biogenic amines. Which foods have you tried that are high in proline but low in amines generated thru protein aging?
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
I'm not sure about amines. Gelatin was the worst offender. Wheat was probably second (very high in proline), with whole wheat having a stronger effect than white (correlating with proline content). Oats had the same effect as wheat, but required more oats for the same severity. Peanut butter and soy butter also correlated with proline content.
 

vision blue

What big eyes...
Messages
1,643
Likes
2,128
Interesting list. Thanks. Gelatin is huge for amines. But not wheat, white or oats as far as i know. Soy butter yes. Probably peanut butter yes too.
How are you with smoked meats, asian fish sauce. I will have to think of stuff high in amines but not in proline. AA tests would provide support for hypothesis.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
How are you with smoked meats, asian fish sauce.
Smoked meats don't seem worse than fresh meat. I never liked fish sauce. Milk and figs (high in proline) are bad for me too.

It's very hard to find foods low in proline and not low in everything else. Chia seeds have some AAs fairly high compared to proline. I'll buy some next time I go to town. I'll also try to get some essential AA capsules, and maybe some taurine or other non-essential AAs.
 

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
New twist: chicken and pork make my symptoms worse, but beef seems to reduce them. Any ideas for how beef differs nutritionally from pork and chicken? My first thought was selenium, but pork supposedly has more than beef, as does whole wheat, so it's probably not that. Any other ideas?

Assuming that it wasn't just a temporary response to the three beef meals I've had so far, I certainly won't complain about beef as a treatment. Such nice, pleasant, ... tasty animals. :D
 
Messages
246
Likes
237
Some weeks ago, I had what I think was a viral infection, and it made my ME symptoms worse after high-protein meals. After the infection was gone, I remained sensitive to proteins. After much experimentation, I determined that I was sensitive to proline. If I consume ~1400 mg of proline, I'll get a noticeable increase in ME symptoms, most noticeably brainfog and muscle aches (I assume neuropathic). Larger doses of proline makes the severity much worse. The time for the decrease in symptoms depends on the dose: just past the threshold may take hours; much larger doses can take days. My response to proline seems a lot like PEM, which in turn seems a lot like an increase in my usual ME symptoms, so this might provide an insight into the core dysfunction of ME and/ or PEM.

Why might proline be involved in ME? Well, proline is critical for mitochondrial function. This paper explains it: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4752965/

"Proline dehydrogenase (PRODH), a.k.a. proline oxidase (POX), is a mitochondrial inner membrane protein. Oxidation of proline to pyrroline-5-carboxylate (P5C) mediates the proline cycle that shuttles NADP(H)/NADP(+) redox equivalents between mitochondria and cytosol and forms a metabolic interlock with the pentose phosphate pathway"

"Proline dehydrogenase/oxidase (PRODH/POX) is a mitochondrial protein critical to multiple stress pathways."

"We found that PRODH/POX binds directly to CoQ1 and that CoQ1-dependent PRODH/POX activity required functional Complex III and Complex IV. PRODH/POX supported respiration in living cells during nutrient stress; however, expression of PRODH/POX resulted in an overall decrease in respiratory fitness. Effects on respiratory fitness were inhibited by DHP and NAC, indicating that these effects were mediated by PRODH/POX-dependent reactive oxygen species (ROS) generation. PRODH/POX expression resulted in a dose-dependent down-regulation of Complexes I–IV of the ETC, and this effect was also mitigated by the addition of DHP and NAC."

To me, it certainly seems like it's possible for proline to be involved in the cellular dysfunction found in ME. I came across a couple of papers that showed that excess proline can reduce mitochondrial function in the brain, an effect that can be reduced by pre-treatment with antioxidants. I'll have to experiment with that to see if it reduces my sensitivity.

Abnormal proline metabolism might explain the collagen problems some PWME report, since proline is critical for collagen formation. Alternatively, maybe a problem with maintaining collagen worsens ME by releasing more proline to muck up mitochondria.

There's a human brain-specific high affinity L-proline transporter (hPROT). "These findings warrant the consideration of a synaptic regulatory role for this transporter and its presumed natural substrate, L-proline, in the mammalian central nervous system." That sounds like it could be one of those biological mechanisms that no one notices when doing ordinary medical testing, but whose subtle dysfunction could result in significant symptoms.

I assume that my present sensitivity to proline is unusual, so I don't think that ME is proline sensitivity. However, this link between proline and ME/PEM symptoms might be worth investigating further, so I'm posting this to see if it triggers some ideas in other people, or makes some researcher evaluate test results from a new perspective.

Also, for people who are wondering why some foods make them feel worse, you could check to see if they're proline-rich. Wheat is very high in protein (there goes another staple from my diet :(). Peanuts are too, and of course, meats. Proline sensitivity is very annoying, diet-wise. :grumpy:


One suggestion for a simple test for researchers: do the nanoneedle test with extra proline added to the fluid. Doing it with extra antioxidants and extra proline might be worthwhile too.
Not sure what is going on, but according to the Kalish institute the ideal range of amino acids in your urine/blood is the highest quartile in the range on a nutreval, ion panel, or similar test. These tests cost anywhere from $400 to $1000, the more expensive ones are ones you can buy online yet the cheaper ones are sold to naturopath or functional medicine practicioners. Interestingly, you can do a blood and urine amino acid profile and use the reference ranges from sample reports of nutreval or ion panel, etc to figure out what a naturopathic practicioner would say if you have too much or too little. You may want to look into Richard Lord, he is considered the grandfather of functional medicine.
 
Last edited by a moderator:

Wishful

Senior Member
Messages
4,766
Likes
8,661
Location
Alberta
Just a followup: right now I feel pretty good, almost a temporary remission. Eating beef may not be an actual long-term treatment, but it sure boosts my spirits to know that this isn't a permanent decline in my health.

I wish all of us could experience temporary improvements to boost our spirits. :hug: