Understand and Test Your Methylation Cycle with a Methylation Panel

Table of Contents

As a quick reminder, methylation is a biochemical process used by the body in processes like regulating neurotransmitters, steroid hormones, inflammation, and DNA expression and producing key compounds such as creatine, phosphatidylcholine, coenzyme Q10, carnitine, melatonin, and epinephrine/adrenaline.

Methylation is simply the addition of a small molecule called a "methyl group" to another molecule. Methyl groups consist of one carbon atom and three hydrogen atoms.

Sometimes methylation occurs in one step, and sometimes it occurs in several steps that add the carbon atom and three hydrogen atoms.

Before describing the Methylation Panel test, let's look in detail at why you might want to use the test, how methylation is used by the body, nutrients and metabolites involved in the Methylation Cycle and the Transsulfuration Pathway, and finally what a methylation panel measures that can provide insight into what might not be working optimally in the Methylation Cycle and/or Transsulfuration Pathway.

Symptoms of Methylation Problems

Here are some common disorders and symptoms of problems with methylation:

Chronic Fatigue Syndrome: fatigue, "brain fog", inability to focus, and more
Low adrenal function: difficulty producing adrenaline from noradrenaline via phenylethanolamide methyltransferase (PNMT)
Histamine intolerance: inability to efficiently clear histamine via histamine N-methyl transferase (HNMT)
Migraines: can be due to inability to efficiently clear histamine
Sleep problems: can be due to difficulty producing melatonin, the "sleep hormone", via acetylserotonin O-methyltransferase (ASMT)
Cancer: usually includes reductions in DNA methylation and corresponding loss of epigenetic regulatory mechanisms
Cardiovascular disease: can involve elevated homocysteine that elevates the risk of cardiovascular disease
Estrogen dominance: reduced libido, reduced energy, heavy or irregular periods, symptoms of low progesterine (even if progesterone is not low) like short cycles and spotting, breast tenderness, fibroids, PMS, headaches and migraines, hair loss (thinning and/or brittle), weight gain around the hips and buttocks, trouble falling asleep and reduction in sleep quality, problems with concepton, bloating, change in bowel movements, water retention in hands and feet, polycystic ovarian disease (PCOS), thyroid disease, and more
Almost any type of neuropsychiatric disorder: anxiety, depression, ADD/ADHD, biopolar disorder, schizophrenia, and more
Megaloblastic macrocytic anemia: fatigue, breathlessness, difficulty walking and vision problems, pain or tingling sensations, confusion, forgetfulness or memory loss, slower thinking, mood changes, unexplained weight loss, diarrhea, changes in smell or taste, and more

Obviously, methylation is involved in many processes.

How Methylation is Used in the Body

The Methylation Cycle produces S-adenosylmethionine (SAM), the main methyl donor in the body, and multiple methyltransferase enzymes transfer the methyl group carried by SAM to other molecules to either synthesize a new biochemically active substance or to inactivate a substance.

Catechol-O-methyltransferase (COMT) methylates dopamine, norepinephrine, epinephrine, steroid hormones like estrogen and cortisol, certain drugs, and quercetin and other flavonoids containing catechol groups. Typically, methylation by COMT inactivates substances in preparation for clearance from the body, but not always. For example, COMT can perform the final step in the conversion of serotonin to melatonin.

Histamine N-methyltransferase (HNMT) methylates histamine as the first step in inactivating histamine.

Phenylethanolamine-N-methyltransferase (PNMT) methylates noradrenaline to convert it to adrenaline.

Acetylserotonin O-methyltransferase (ASMT) can perform the final step in the conversion of serotonin to melatonin. COMT can also perform this step.

Phosphatidylethanolamine N-methyltransferase (PEMT) transfers three methyl groups to phosphatidylethanolamine to form phosphatidylcholine (PC), the precursor for the neurotransmitter acetylcholine and an essential component of cell membranes., bile, and lipoproteins that transport cholesterol and other lipids. .

Guanidinoacetate N-methyltransferase (GAMT) performs the last step in the synthesis of creatine, which helps the muscles and brain perform better by quickly recycling ADP back to ATP under conditions of high energy demand.

DNA methyltransferases (DNMT) methylate DNA to turn off the expression of genes as one of the primary mechanisms of epigenetic regulation.

And there are other methyltransferase enzymes not described above.

The Methylation Cycle and the Transsulfuration Pathway

In the health space, methylation is often oversimplified to discussions and testing of single nucleotide polymorphisms (SNPs) in the MTHFR gene/enzyme. But so much more is involved in the Methylation Cycle than MTHFR, as you will see in the next section.

The Methylation Cycle also interfaces with a pathway called the Transsulfuration Pathway, which must be considered in any discussion of the Methylation Cycle.

Transsulfuration is another way that the body metabolizes homocysteine, the inflammatory metabolite in the Methylation Cycle that the Methylation Cycle converts back to methionine.

The Transsulfuration Pathway converts homocysteine to cysteine, the "rate-limiting" amino acid required for the production of glutathione, the body's "master antioxidant".

In order to truly understand these pathways, we need to look at all of, or at least most of, the enzymes / genes involved and the nutrients these enzymes require for their function.

Roles of Key Enzymes/Genes and Nutrients in Methylation and Transsulfuration

There are at least nine key enzymes / genes involved in the Methylation Cycle, including DHFR, SHMT, MTHFD1, MTHFR, MTR, MTRR, BHMT, MAT, and AHCY.

And there are three key enzymes / genes involved in the Transsulfuration Cycle, including CBS, CTH, and SUOX.

Most of the enzymes in the Methylation Cycle and the Transsulfuration Pathway require certain nutrients as cofactors in order to function.

DHFR (dihydrofolate reductase) converts folic acid (synthetic) to diydrofolate (DHF) and converts DHF (also from dietary sources) to tetrahydrofolate (THF).

MTHFD (methylenetetrahydrofolatedehydrogenase) converts THF to 5,10-methylene-THF in several steps, using dimethylglycine (DMG) or sarcosine as the methyl donor for the methylation step. MTHFD can also perform other roles in the folate cycle.

SHMT (serine hydroxymethyltransferase) transfers a methyl group from serine to convert THF to 5, 10-methylene-THF in on step. SHMT requires B6 as its cofactor.

MTHFR (methylenetetrahydrofolate reductase) transfers an electron from NADPH to convert 5, 10-methylene-THF to 5, 10-methyl-THF. MTHFR requires vitamin B2 as the precursor for its coenzyme, flavin adenine dinucleotide (FAD).

MTR (methionine synthase) transfers a methyl group from methylfolate (via methyl-B12 and the MTRR enzyme) to homocysteine to convert it to methionine. It requires MTRR for the part of the process that involves remethylating methyl-B12. MTR requires zinc as its cofactor.

MTRR (methionine synthase reductase) works with MTR to transfer the methyl group from methylfolate to homocysteine to convert it to methyionine. MTRR also recycles oxidized B12. It requires vitamin B2 as the precursor for its coenzymes, flavin adenine dinucleotide (FAD) and flavin mononucleotide (FMN).

BHMT (betaine-homocysteine methyltransferase) converts homocysteine to methionine by using a methyl group from betaine / trimethylglycine (TMG), which is derived from choline. BHMT requires zinc as its cofactor.

MAT (methionine adenosyltransferase / S-adenosyl methionine synthetase) converts methionine to S-adenosyl-methionine (SAM/SAMe), the body's "universal methyl donor". MAT use adenosine triphosphate (ATP) as the source of the adenosyl group that it transfers to methionine to produce SAM. MAT is particularly vulnerable to the hydroxyl radical.

Methyl transferase enzymes like COMT, HNMT, PNMT, and ASMT transfer a methyl group from SAM to other molecules, causing SAM to become S-adenosyl-homocysteine (SAH). COMT requires magnesium as its cofactor.

AHCY converts SAH to homocysteine. It uses an electron from NADPH in the process.

CBS converts homocysteine to cystathionine, using a methyl group from serine. It requires vitamin B6 for its function.

CTH converts cystathionine to cysteine, which can then be used to produce glutahtione.

If CBS and is highly active, it can lead to elevated production of sulfite, which is neurotoxic in excess.

SUOX converts neurotoxic sulfide to sulfate, which is used in numerous sulfation processes such phase II of biotransformation, which most of us know as "detoxification".

Many of these enzymes *e.g. MTHFD, MTHFR and MTRR) requjre require NADPH as an electron donor, which can be depleted in many types ofillness.

Anywhere you see "B3" as a cofactor in the images below from a Genova Diagnostics Methylation Panel depicting the Methylation Cycle and the Transsulfuration Pathway, it is an indication of where NADPH is required.

Methylation Cycle and Transsulfuration Pathway

The image above is from a Genova Diagnostics Methylation Panel, the best "metabolomic" methylation panel I know of.

Metabolomic vs Genomic Methylation Panels

Some test providers use the term "methylation panel" to refer to a genomic test of the single nucleotide polymorphisms (SNPs) in the genes involved in the Methylation Cycle, such as MTHFR, MTHFD, SHMT, BHMT, MTR, MTRR, etc.

But a genomic test only tells you the likelihood of developing problems in various parts of the methylation cycle. It does not tell you where problems might be showing up today in a one's biochemistry, which could be quite different.

I recommend using a metabolomic methylation panel that tests certain nutrients and key metabolites produced in the methylation cycle, whether or not you also use a genetic methylation panel.

The most complete methylation panel I know of is the Methylation Panel provided by Genova Diagnostics.

It tests the following metabolites and nutrients:

S-adenosyl-methionine (SAM)
S-adenosyl-homocysteine (SAH)
Homocysteine
Cystathionine
Glutathione
Choline
Betaine / trimethylglycine (TMG)
Dimethylglycine (DMG)
Sarcosine
Methionine
Serine
Glycine
Cysteine
Taurine

The image below is from a Genova Diagnostics Methylation Panel.

If any of these markers are high or low, it provides clues as to which nutrients may be deficient and which steps in the Methylation Cycle or Transsulfuration Pathway may be hindered (or overactive in the case of the CBS enzyme and elevated cystathionine).

Genova Diagnostics has an option to test methylation genes as an add-on feature of the Methylation Panel, but I recommend using whole genome sequencing as a much more cost-effective way to learn about these genes and all of your other genes.

The best way I know of to truly understand what is happening in a person's methylation cycle is to run a metabolomic methylation panel, a genomic methylation panel (or full genome sequencing), an organic acids test, and a test of B vitamins and certain minerals in serum and cells.

However, if you cannot get all of these tests, the metabolomic Methylation Panel could be enough to troubleshoot and resolve methylation challenges.

Reasons to Get a Methylation Panel

Because the Methylation Cycle is somewhat complex, knowing a few things like the status of folate, B12, and MTHFR SNPs often is not enough to solve problems with methylation.

If you need to get to the bottom of a health challenge that could involve methylation, full methylation testing, including a metabolomic Methylation Panel, can reveal important clues about what may have been keeping you stuck.

Some of the conditions and symptoms (repeated from earlier in this article) commonly linked to problems in the Methylation Cycle are listed below:

Chronic Fatigue Syndrome: fatigue, "brain fog", inability to focus, and more
Low adrenal function: difficulty producing adrenaline from noradrenaline via phenylethanolamide methyltransferase (PNMT)
Histamine intolerance: inability to efficiently clear histamine via histamine N-methyl transferase (HNMT)
Migraines: can be due to inability to efficiently clear histamine
Sleep problems: can be due to difficulty producing melatonin, the "sleep hormone", via acetylserotonin O-methyltransferase (ASMT)
Cancer: usually includes reductions in DNA methylation and corresponding loss of epigenetic regulatory mechanisms
Cardiovascular disease: can involve elevated homocysteine that elevates the risk of cardiovascular disease
Estrogen dominance: reduced libido, reduced energy, heavy or irregular periods, symptoms of low progesterine (even if progesterone is not low) like short cycles and spotting, breast tenderness, fibroids, PMS, headaches and migraines, hair loss (thinning and/or brittle), weight gain around the hips and buttocks, trouble falling asleep and reduction in sleep quality, problems with concepton, bloating, change in bowel movements, water retention in hands and feet, polycystic ovarian disease (PCOS), thyroid disease, and more
Almost any type of neuropsychiatric disorder: anxiety, depression, ADD/ADHD, biopolar disorder, schizophrenia, and more
Megaloblastic macrocytic anemia: fatigue, breathlessness, difficulty walking and vision problems, pain or tingling sensations, confusion, forgetfulness or memory loss, slower thinking, mood changes, unexplained weight loss, diarrhea, changes in smell or taste, and more

If you would like to discuss whether a Methylation Panel and/or other tests might serve you well as next steps on your health journey, you can schedule a free 15-minute consultation with me to discuss what you're looking to accomplish, what you've done so far, and some suggestions about testing to consider.

To learn about my approach to resolving health challenges and optimizing energy, sleep, immunity, and more, download the Bio-Individual Blueprint Roadmap and watch the walkthrough video.