Presence of thiamine pyrophosphate in mammalian peroxisomes

Department of Molecular Cell Biology, Division of Pharmacology, LIPIT, Katholieke Universiteit Leuven, O&N1, Leuven, Belgium. <>
BMC Biochemistry (Impact Factor: 1.94). 02/2007; 8:10. DOI: 10.1186/1471-2091-8-10
Source: PubMed

ABSTRACT Thiamine pyrophosphate (TPP) is a cofactor for 2-hydroxyacyl-CoA lyase 1 (HACL1), a peroxisomal enzyme essential for the alpha-oxidation of phytanic acid and 2-hydroxy straight chain fatty acids. So far, HACL1 is the only known peroxisomal TPP-dependent enzyme in mammals. Little is known about the transport of metabolites and cofactors across the peroxisomal membrane and no peroxisomal thiamine or TPP carrier has been identified in mammals yet. This study was undertaken to get a better insight into these issues and to shed light on the role of TPP in peroxisomal metabolism.
Because of the crucial role of the cofactor TPP, we reanalyzed its subcellular localization in rat liver. In addition to the known mitochondrial and cytosolic pools, we demonstrated, for the first time, that peroxisomes contain TPP (177 +/- 2 pmol/mg protein). Subsequently, we verified whether TPP could be synthesized from its precursor thiamine, in situ, by a peroxisomal thiamine pyrophosphokinase (TPK). However, TPK activity was exclusively recovered in the cytosol.
Our results clearly indicate that mammalian peroxisomes do contain TPP but that no pyrophosphorylation of thiamine occurs in these organelles, implying that thiamine must enter the peroxisome already pyrophosphorylated. Consequently, TPP entry may depend on a specific transport system or, in a bound form, on HACL1 translocation.

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Available from: Minne Casteels, Dec 25, 2013
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    • "Three replicates of each sample were extracted in 500 µl of 7.2% perchloric acid by sonicating for 30 min in a bath, held on ice for 15 min with periodic vortex mixing, and then cleared by centrifugation at (14000 g, 10 min). The supernatant was then analyzed for thiamin and its phosphates by oxidation to thiochrome derivatives followed by HPLC with fluorometric detection (Fraccascia et al., 2007; Goyer et al., 2013). "
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    ABSTRACT: The B vitamin thiamin is essential for central metabolism in all cellular organisms including plants. While plants synthesize thiamin de novo, organs vary widely in their capacities for thiamin synthesis. We use a transcriptomics approach to appraise the distribution of de novo synthesis and thiamin salvage pathways among organs of maize. We identify at least six developmental contexts in which metabolically active, non-photosynthetic organs exhibit low expression of one or both branches of the de novo thiamin biosynthetic pathway indicating a dependence on inter-cellular transport of thiamin and/or thiamin precursors. Neither the thiazole (THI4) nor pyrimidine (THIC) branches of the pathway are expressed in developing pollen implying a dependence on import of thiamin from surrounding floral and inflorescence organs. Consistent with that hypothesis, organs of the male inflorescence and flowers are shown to have high relative expression of the thiamin biosynthetic pathway and comparatively high thiamin contents. By contrast, divergent patterns of THIC and THI4 expression occur in the shoot apical meristem, embyro sac, embryo, endosperm, and root-tips suggesting that these sink organs acquire significant amounts of thiamin via salvage pathways. In the root and shoot meristems, expression of THIC in the absence of THI4 indicates a capacity for thiamin synthesis via salvage of thiazole, whereas the opposite pattern obtains in embryo and endosperm implying that seed storage organs are poised for pyrimidine salvage. Finally, stable isotope labeling experiments set an upper limit on the rate of de novo thiamin biosynthesis in maize leaf explants. Overall, the observed patterns of thiamin biosynthetic gene expression mirror the strategies for thiamin acquisition that have evolved in bacteria.
    Frontiers in Plant Science 08/2014; 5:370. DOI:10.3389/fpls.2014.00370 · 3.95 Impact Factor
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    • "In addition, TPP is involved in the í µí»¼oxidation of 3-methyl-branched and straight chain 2-hydroxy long chain fatty acids pathway functioning as coenzyme for peroxisomes. As a result TPP is a crucial cofactor for energy metabolism, antioxidation, and myelinization of nerve cells [21]. Continued high-dose cisplatin chemotherapy necessitates the investigation of strategies to decrease the doselimiting ototoxicity. "
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    The Scientific World Journal 09/2013; 2013:182694. DOI:10.1155/2013/182694 · 1.73 Impact Factor
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    • "The first allusion to the existence of thiamine-binding sites on biological membranes, different from intra membrane carrier, was suggested trough the reversal of the effects of thiamine deficiency by antioxidants, suggesting that thiamine may act as a site-directed antioxidant (Gibson and Zhang 2002). Recent disclosure in peroxisomes of TDP-dependent enzyme that catalyzes alpha-oxidation of straight chain fatty acids makes possible the existence of membrane surface sites fixing thiamine (Sniekers et al. 2006; Fraccascia et al. 2007). Through our developmental studies, we reported that the specific need of B1 vitamin increased in nervous tissues during ontogenesis and corresponded to the post-natal development of membrane processes (Bâ 2005). "
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    Cellular and Molecular Neurobiology 11/2008; 28(7):923-31. DOI:10.1007/s10571-008-9297-7 · 2.20 Impact Factor
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