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Dietary methyl donors affect in vivo methionine partitioning between transmethylation and protein synthesis in the neonatal piglet

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Abstract

Methionine metabolism is critical during development with significant requirements for protein synthesis and transmethylation reactions. However, separate requirements of methionine for protein synthesis and transmethylation are difficult to define because after transmethylation, demethylated methionine is either irreversibly oxidized to cysteine during transsulfuration, or methionine is regenerated by the dietary methyl donors, choline (via betaine) or folate during remethylation. We hypothesized that remethylation contributes significantly to methionine availability and affects partitioning between protein and transmethylation. 4–8-day-old neonatal piglets were fed a diet devoid (MD−) (n = 8) or replete (MS+) (n = 8) of folate, choline and betaine to limit remethylation. After 5 days, dietary methionine was reduced to 80 % of requirement in both groups of piglets to ensure methionine availability was limited. On day 7, an intragastric infusion of [13C1]methionine and [2H3-methyl]methionine was administered to measure methionine cycle flux. In MD− piglets, in vivo remethylation was 60 % lower despite 23-fold greater conversion of choline to betaine (P < 0.05) and transmethylation was 56 % lower (P < 0.05), suggesting dietary methyl donors spared 425 µmol methyl/day for transmethylation. The priority of protein synthesis versus transmethylation was clear during MD− feeding (P < 0.05), as an additional 6 % of methionine flux was for protein synthesis in those piglets (P < 0.05). However, whole body transsulfuration was unaffected in vivo despite reduced in vitro cystathionine-β-synthase capacity in MD− piglets (P < 0.05). Our data show that remethylation contributes significantly to methionine availability and that transmethylation is sacrificed to maintain protein synthesis when methionine is limiting in neonates, which should be considered when determining the methionine requirement.

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Abbreviations

APE:

Atom percent excess

BHMT:

Betaine:homocysteine methyltransferase

CBS:

Cystathionine-β-synthase

DMG:

Dimethylglycine

FNC:

Fractional net conversion

FPF:

Fraction of product flux

MSyn:

Methionine synthase

Q :

Flux

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Authors and Affiliations

  1. Department of Biochemistry, Memorial University of Newfoundland, St. John’s, NL, A1B 3X9, Canada

    Jason L. Robinson, Renee K. Bartlett, Janet A. Brunton & Robert F. Bertolo

  2. Diabetes and Nutritional Sciences Division, Faculty of Life Sciences and Medicine, King’s College London, London, SE1 9NH, UK

    Scott V. Harding

  3. Department of Laboratory Medicine, Faculty of Medicine, Memorial University of Newfoundland, St. John’s, NL, A1B 3X9, Canada

    Edward W. Randell

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  1. Jason L. Robinson
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  2. Renee K. Bartlett
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  4. Edward W. Randell
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  5. Janet A. Brunton
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  6. Robert F. Bertolo
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Correspondence to Robert F. Bertolo.

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Funding disclosure

This project was funded with support from the Canadian Institutes of Health Research (Grant number 201103RNL) and the Research & Development Corporation of Newfoundland and Labrador (Grant number 5404-1046-104) awarded to RFB. JLR was supported by a fellowship from the Canadian Institutes of Health Research and the Research & Development Corporation of Newfoundland and Labrador.

Conflict of interest

JLR, LEM, EWR, JAB, RFB declare no conflicts of interest.

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Handling Editor: C.-A. A. Hu.

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Robinson, J.L., Bartlett, R.K., Harding, S.V. et al. Dietary methyl donors affect in vivo methionine partitioning between transmethylation and protein synthesis in the neonatal piglet. Amino Acids 48, 2821–2830 (2016). https://doi.org/10.1007/s00726-016-2317-x

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