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Molecules 2016, 21(9), 1195; doi:10.3390/molecules21091195

Comparison of Protein N-Homocysteinylation in Rat Plasma under Elevated Homocysteine Using a Specific Chemical Labeling Method

1
Barnett Institute of Chemical and Biological Analysis, Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115, USA
2
Human Nutrition Research Center on Aging, Tufts University, Boston, MA 02111, USA
3
Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, MA 02115, USA
4
Department of Pharmaceutical Sciences, Nova Southeastern University, Fort Lauderdale, FL 33314, USA
5
School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou 325035, China
*
Authors to whom correspondence should be addressed.
Academic Editor: Derek J. McPhee
Received: 16 August 2016 / Revised: 2 September 2016 / Accepted: 5 September 2016 / Published: 8 September 2016
(This article belongs to the Section Bioorganic Chemistry)
View Full-Text   |   Download PDF [2443 KB, uploaded 8 September 2016]   |  

Abstract

Elevated blood concentrations of homocysteine have been well established as a risk factor for cardiovascular diseases and neuropsychiatric diseases, yet the etiologic relationship of homocysteine to these disorders remains poorly understood. Protein N-homocysteinylation has been hypothesized as a contributing factor; however, it has not been examined globally owing to the lack of suitable detection methods. We recently developed a selective chemical method to label N-homocysteinylated proteins with a biotin-aldehyde tag followed by Western blotting analysis, which was further optimized in this study. We then investigated the variation of protein N-homocysteinylation in plasma from rats on a vitamin B12 deficient diet. Elevated “total homocysteine” concentrations were determined in rats with a vitamin B12 deficient diet. Correspondingly, overall levels of plasma protein N-homocysteinylation displayed an increased trend, and furthermore, more pronounced and statistically significant changes (e.g., 1.8-fold, p-value: 0.03) were observed for some individual protein bands. Our results suggest that, as expected, a general metabolic correlation exists between “total homocysteine” and N-homocysteinylation, although other factors are involved in homocysteine/homocysteine thiolactone metabolism, such as the transsulfuration of homocysteine by cystathionine β-synthase or the hydrolysis of homocysteine thiolactone by paraoxonase 1 (PON1), may play more significant or direct roles in determining the level of N-homocysteinylation. View Full-Text
Keywords: hyperhomocysteinemia; cardiovascular disease; neuropsychiatric disease; protein N-homocysteinylation; plasma; biotin-aldehyde; Western blotting hyperhomocysteinemia; cardiovascular disease; neuropsychiatric disease; protein N-homocysteinylation; plasma; biotin-aldehyde; Western blotting
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Zang, T.; Pottenplackel, L.P.; Handy, D.E.; Loscalzo, J.; Dai, S.; Deth, R.C.; Zhou, Z.S.; Ma, J. Comparison of Protein N-Homocysteinylation in Rat Plasma under Elevated Homocysteine Using a Specific Chemical Labeling Method. Molecules 2016, 21, 1195.

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