Glutathione depletion and disruption of the sleep-wake cycle in ME/CFS

Discussion in 'Fibromyalgia Main Forum' started by richvank, Aug 12, 2011.

  1. richvank

    richvank New Member

    Hi, all.

    As many of you know, I am the proponent of the Glutathione
    Depletion--Methylation Cycle Block hypothesis for the pathogenesis and
    pathophysiology of ME/CFS.

    If glutathione depletion coupled with a partial block in the methylation
    cycle is indeed the core mechanism in the pathophysiology of ME/CFS, then the
    various abnormalities in ME/CFS must stem from it. In the past, I have been able
    to find biochemical connections between this basic mechanism and many of the
    abnormalities in ME/CFS.

    One feature of ME/CFS that I have not yet connected to this mechanism is the
    disruption in the sleep-wake cycle that many PWMEs experience.

    Recently I came upon a paper that I think will enable me to make this
    connection. The abstract is below, and the full paper is available from PubMed.

    This paper discusses the presence of a disulfide bridge in the serotonin
    N-acetyltransferase enzyme molecule. This enzyme catalyzes one of the steps in
    the conversion of serotonin to melatonin, and this conversion is important in
    controlling the sleep-wake cycle. Under reducing conditions, the disulfide
    bridge does not form, and the enzyme can catalyze its reaction. Under oxidizing
    conditions, when the disulfide bridge forms, the reaction is not catalyzed.

    I suggest that if glutathione becomes depleted in the pineal gland, the
    resulting oxidizing conditions will cause formation of this disulfide bond,
    shutting off the conversion of serotonin to melatonin, and disrupting the
    sleep-wake cycle.

    If this is true, I think we can expect that if glutathione is brought back
    up by lifting the partial methylation cycle block using one of the methylation
    treatment protocols, the sleep-wake cycle abnormalities in ME/CFS should be
    corrected. I note that some people who have tried this type of treatment (including those in the clinical trial conducted by Neil Nathan, M.D., have
    indeed reported improvements in sleep.

    Best regards,


    J Biol Chem. 2002 Nov 15;277(46):44229-35. Epub 2002 Sep 4.

    An intramolecular disulfide bridge as a catalytic switch for serotonin

    Tsuboi S, Kotani Y, Ogawa K, Hatanaka T, Yatsushiro S, Otsuka M, Moriyama Y.

    Department of Biochemistry, Faculty of Pharmaceutical Sciences, Okayama
    University, Japan.

    Serotonin N-acetyltransferase (EC. (AA-NAT) is a melatonin
    rhythm-generating enzyme in pineal glands. To establish a melatonin rhythm,
    AA-NAT activity is precisely regulated through several signaling pathways. Here
    we show novel regulation of AA-NAT activity, in which an intramolecular
    disulfide bond may function as a switch for the catalysis. Recombinant AA-NAT
    activity was irreversibly inhibited by N-ethylmaleimide (NEM) in an
    acetyl-CoA-protected manner. Oxidized glutathione or dissolved oxygen reversibly
    inhibited AA-NAT in an acetyl-CoA-protected manner. To identify the cysteine
    residues responsible for the inhibition, AA-NAT was first oxidized with
    dissolved oxygen, treated with NEM, reduced with dithiothreitol, and then
    labeled with [(14)C]NEM. Cys(61) and Cys(177) were specifically labeled in an
    acetyl-CoA-protected manner. The AA-NAT with the Cys(61) to Ala and Cys(177) to
    Ala double substitutions (C61A/C177A-AA-NAT) was fully active but did not
    exhibit sensitivity to either oxidation or NEM, whereas the AA-NATs with only
    the single substitutions retained about 40% of these sensitivities. An
    intramolecular disulfide bond between Cys(61) and Cys(177) formed upon oxidation
    and cleaved upon reduction was identified. Furthermore, C61A/C177A-AA-NAT
    expressed in COS7 cells was relatively insensitive to H(2)O(2)-evoked oxidative
    stress, whereas wild-type AA-NAT was strongly inhibited under the same
    conditions. These results indicate that the formation and cleavage of the
    disulfide bond between Cys(61) and Cys(177) produce the active and inactive
    states of AA-NAT. It is possible that intracellular redox conditions regulate
    AA-NAT activity through switching via an intramolecular disulfide bridge.


    (Please note that this abstract is from PubMed, which is operated by NIH, part of the U.S. government. Abstracts on PubMed are in the public domain with respect to copyright.)[This Message was Edited on 08/12/2011]
  2. Scapper

    Scapper New Member

    Thank you for this informative article Rich!

    Improved sleep would be WONDERFUL!!! It's nice to even hear that it's a possibility.

    I soooooo appreciate all that you bring here!!!!!!!!!!!!

  3. Mikie

    Mikie Moderator

    Thanks for the info and for pointing out that it's in the public domain.

    Love, Mikie
  4. mbofov

    mbofov Active Member

    Thanks again - more fascinating information!

    But what I am still curious about is, what causes the methylation block in the first place, particularly in people who have been taking B vitamins and B12 for many years? (unless the culprit is folic acid?)

  5. richvank

    richvank New Member

    Hi, Mary,

    In my hypothesis, ME/CFS onset occurs when stressors of various types (physical, chemical, biological and psychological/emotional) contribute to depleting glutathione in a person who is genetically susceptible.

    When glutathione goes low enough, it is not possible for the cells to make methyl B12 from the various forms of B12 that are coming in from the diet (and supplements, if they are also being taken). This produces a functional deficiency in B12, rather than an absolute deficiency. In other words, the person may be taking in enough B12, but their cells are not able to make proper use of it because of the glutathione depletion.

    The lack of sufficient methyl B12 then causes an inhibition of the activity of methionine synthase in the methylation cycle, producing a partial block. That in turn causes methylfolate to drain from the cells to the blood stream. The partial block of methionine synthase stabilizes the glutathione depletion in a vicious circle mechanism, and that makes ME/CFS a chronic condition.

    In order to overcome this vicious circle, it is necessary to take a much larger dosage of B12 than is usually needed by a healthy person, together with a dosage of active folate that is comparable to normal RDA levels. If there are deficiencies in cofactor vitamins and minerals or in necessary amino acids, these need to be supplemented as well.

    Best regards,

    [This Message was Edited on 08/14/2011]
  6. munch1958

    munch1958 Member

    I've been using nebulized glut for MCS since the fall of 2006. I don't have hardly any symptoms as long as I keep doing it 3x a week and stick with my hormones especially hydrocortisone. If I fall behind for any reason then the world gets a little more stinky and smells start to bother me. I also have learned that if I'm going to be in heavy city traffic in Chicago as opposed to my rural area to double up my dose the night before.

    I can say without a doubt that on the days I nebulize glut: I sleep like a rock. Since I do it 3 days a week, there is a bit of a gap for 2 days of the week and that is when I have trouble falling and staying asleep.

  7. richvank

    richvank New Member

    Hi, Munch.

    I'm glad that you get relief from MCS by using nebulized glutathione. I think it would be interesting to know if you have a partial methylation cycle block. If you do, it may be possible to lift it and thereby raise glutathione on a permanent basis, so you wouldn't have to continue nebulizing it. If you are interested in finding out about testing for a partial methylation cycle block, you can find my papers at

    The most recent version of the simplified treatment protocol is there, too.

    Best regards,

  8. mbofov

    mbofov Active Member

    I know you've explained this to us a dozen times, but thanks for doing it again!

    But my question is this - the protocol is essentially B12 and folate, with vitamins, amino acids, and minerals as needed. My doctor had me doing large doses of B12 for many years. Sometimes I would do hydroxocobalamin, and sometimes I would do methylcobalamin shots, but in any event, a rather large dose - 5,000 mcg. 3 x a week (self-administered). In addition, I've taken B complex vitamins (plus other vitamins and minerals and amino acids and fish oil, etc.), which always included folic acid, for many years.

    So I'm wondering, if the B complex I'd been taking all these years had had an active form of folate instead of folic acid, perhaps I would not be suffering from this methylation block? Was the folic acid a major contributor to the methylation block? (also because of its propensity for competing for absorption with folate)

    Having said all this, I've just been reading Vegas' posts on the PR board, about chelation. I am doing better on Freddd's protocol, but I still crash just as often. Vegas said she overcame PEM with chelation using Andrew Cutler's protocol, so this is next on list of things to do. I am really tired of all this, but seem to have no choice if I don't want to resign myself to basically sitting on the couch for the rest of my life. I had several mercury fillings (can't remember how many, probably 8 or 10) removed 10 or 12 years ago, but I never did chelation.

    I saw where you posted that someone else did well after doing chelation, after doing teh SMP - I have some relatives coming to visit in about 3 weeks so won't be doing anything until after that.

    Thanks as always -

  9. Kiwijack

    Kiwijack Member

    My sleep has been overwhelmingly non restorative and I wake in significant pain, joints, muscles, bones, feels like every cell is screaming out, a hangover x 100, and this has been happening over a dozen years. If I lay there and meditate sometimes the pain goes away and I can doze off again, but a few minutes later the full force of the pain returns. I obviously sleep, 4-6 hrs ea night, so it feels like a sleep/wake transition issue, cortisol or adrenalin surge, shocks me awake, battle stations, immediately I'm fully awake, don't feel rested at all, just feel sore and exhausted. Afternoon nap is the same thing, but the nap only lasts a minute or 2 and I'm shocked back awake in full pain. Being treated for methylation now, lots of gene defects. This article gives me hope. I need a reset button!