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J Nutr Biochem. 2015 Apr;26(4):312-8. doi: 10.1016/j.jnutbio.2014.10.004. Epub 2014 Nov 12.

The application of a chemical determination of N-homocysteinylation levels in developing mouse embryos: implication for folate responsive birth defects.

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Department of Molecular Biosciences, University of Texas, Austin, USA.
Proteomics Facility, Institute for Cellular and Molecular Biology, and College of Pharmacy, The University of Texas at Austin, Austin, TX, USA.
Department of Nutritional Sciences, Dell Pediatric Research Institute, University of Texas, Austin, USA. Electronic address:


Elevated homocysteine levels have long been associated with various disease states, including cardiovascular disease and birth defects, including neural tube defects (NTDs). One hypothesis regarding the strong correlation between these various disorders and high levels of homocysteine is that a reactive form of this small molecule can attach to mammalian proteins in a phenomenon known as homocysteinylation. These posttranslational modifications may become antigenic or may even directly disrupt certain protein function. It remains to be determined whether dietary influences that can cause globally increased levels of circulating homocysteine confer negative effects maternally, or may otherwise negatively and materially impact the metabolic balance in developing embryos. Herein we present the application of a chemical method of determination of N-homocysteinylation to a set of neural tube closure stage mouse embryos and their mothers. We explore the uses of this newly described technique to investigate levels of maternal and embryonic N-homocysteinylation using dietary manipulations of one-carbon metabolism with two known folate-responsive NTD mouse models. The data presented reveal that although diet appeared to have significant effects on the maternal metabolic status, those effects did not directly correlate to the embryonic folate or N-homocysteinylation status. Our studies indicate that maternal diet and embryonic genotype most significantly affected the embryonic developmental outcome.


Homocysteine; Neural tube defects; One-carbon metabolism; Post-translational modifications; in utero metabolism

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