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Metabolism and neurotoxicity of homocysteine thiolactone in mice: protective role of bleomycin hydrolase

Abstract

Genetic or nutritional disorders in homocysteine (Hcy) metabolism elevate Hcy-thiolactone and cause heart and brain diseases. Hcy-thiolactone has been implicated in these diseases because it has the ability to modify protein lysine residues and generate toxic N-Hcy-proteins with auto-immunogenic, pro-thrombotic, and amyloidogenic properties. Bleomycin hydrolase (Blmh) has the ability to hydrolyze l-Hcy-thiolactone (but not d-Hcy-thiolactone) to Hcy in vitro, but whether this reflects a physiological function has been unknown. Here, we show that Blmh −/− mice excreted in urine 1.8-fold more Hcy-thiolactone than wild-type Blmh +/+ animals (P = 0.02). Hcy-thiolactone was elevated 2.3-fold in brains (P = 0.004) and 2.0-fold in kidneys (P = 0.047) of Blmh −/− mice relative to Blmh +/+ animals. Plasma N-Hcy-protein was elevated in Blmh −/− mice fed a normal (2.3-fold, P < 0.001) or hyperhomocysteinemic diet (1.5-fold, P < 0.001), compared with Blmh +/+ animals. More intraperitoneally injected l-Hcy-thiolactone was recovered in plasma in Blmh −/− mice than in wild-type Blmh +/+ animals (83.1 vs. 39.3 μM, P < 0.0001). In Blmh +/+ mice injected intraperitoneally with d-Hcy-thiolactone, d,l-Hcy-thiolactone, or l-Hcy-thiolactone, 88, 47, or 6.3%, respectively, of the injected dose was recovered in plasma. The incidence of seizures induced by l-Hcy-thiolactone injections (3,700 nmol/g body weight) was higher in Blmh −/− than in Blmh +/+ mice (93.8 vs. 29.5%, P < 0.001). Using the Blmh null mice, we provide the first direct evidence that a specific Hcy metabolite, Hcy-thiolactone, rather than Hcy itself, is neurotoxic in vivo. Taken together, our findings indicate that Blmh protects mice against l-Hcy-thiolactone toxicity by metabolizing it to Hcy and suggest a mechanism by which Blmh might protect against neurodegeneration associated with hyperhomocysteinemia and Alzheimer’s disease.

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Abbreviations

Blmh:

Bleomycin hydrolase

Cbs:

Cystathionine β-synthase

Hcy:

Homocysteine

tHcy:

Total Hcy

i.p.:

Intraperitoneal

Mthfr:

Methylenetetrahydrofolate reductase

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Acknowledgments

We thank John Lazo for kindly providing Blmh null mice and for his comments on the manuscript. This work was supported in part by grants from the American Heart Association (0855919D) and the National Science Center, Poland (DEC-2011/01/B/NZ1/03417).

Conflict of interest

The authors declare that they have no conflict of interest.

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Correspondence to Hieronim Jakubowski.

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Borowczyk, K., Tisończyk, J. & Jakubowski, H. Metabolism and neurotoxicity of homocysteine thiolactone in mice: protective role of bleomycin hydrolase. Amino Acids 43, 1339–1348 (2012). https://doi.org/10.1007/s00726-011-1207-5

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Keywords

  • Bleomycin hydrolase
  • Homocysteine thiolactone
  • Brain
  • Seizures
  • Alzheimer’s disease