Reversed-phase high-performance liquid chromatographic analysis of thiamine phosphate esters at subpicomole levels

Sanemori, Hiroshi; Ueki, Hiroaki; Kawasaki, Takashi

doi:10.1016/0003-2697(80)90409-1

Cited by 39 publications

(5 citation statements)

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“…The two ThTPases were similar to their rat tissue counterparts. Key Words thiamin triphosphate, thiamin triphosphatase, thiamin di phosphate kinase, thiamin pyrophosphate, thiamin, E. coli Recently, data on thiamin triphosphate (ThTP) in the tissues of higher organisms as determined by high performance liquid chromatography (HPLC) have been accumulating (1,2). Those recent reports have reconfirmed earlier discoveries of ThTP in rat tissues (3,4), in yeast (5) and in plant cells (6).…”

Section: Hydrolysis and Synthesis Of Thiamin Triphosphate In Bacteriasupporting

confidence: 62%

Hydrolysis and synthesis of thiamin triphosphate in bacteria.

Nishimune

Hayashi

1987

Journal of Nutritional Science and Vitaminology, J Nutr Sci Vit

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SummaryThiamin triphosphate (ThTP) in early stationary phase cells of Escherichia coli grown in nutrient broth with 0.1% yeast extract was fo und to constitute approximately 5-7% of cellular thiamin diphosphate (ThDP) or around 5nmol/g cell. Nearly the same level of. ThTP was obtained in a Bacillus strain. When E. coli was loaded with an excess of ThTP or ThDP, cellular ThTP was found to be controlled in the course of the long term to maintain its ratio to the amount of cellular ThDP. The ThTP vs. ThDP ratio in E. coli cells after short-term ThDP uptake was fo und to be a function of the cellular growth phase. The ratio in early exponential phase E. coli cells was found to be approximately 4% and it became lower (less than 3%) when cell growth proceeded to the late exponential stage. Two phosphatases specific for ThTP (ThTPase) among thiamin phosphates were detected in E. coli. One required Mgt2+and was fo und mainly in the soluble fraction, while the other was Mgt+ independent and originated from the membrane. The two ThTPases were similar to their rat tissue counterparts. Key Words thiamin triphosphate, thiamin triphosphatase, thiamin di phosphate kinase, thiamin pyrophosphate, thiamin, E. coli Recently, data on thiamin triphosphate (ThTP) in the tissues of higher organisms as determined by high performance liquid chromatography (HPLC) have been accumulating (1, 2). Those recent reports have reconfirmed earlier discoveries of ThTP in rat tissues (3, 4), in yeast (5) and in plant cells (6). In spite of the detections of a small but consistent amount of ThTP, including an exceptionally high percentage of ThTP among the thiamin phosphates in an electric organ of Torpedo marmorata (7) and pig muscle (8), its biological significance has yet to be established. Recent studies on ThTP have mainly focussed on its role in the nervous * This study was carried out at

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Section: Hydrolysis and Synthesis Of Thiamin Triphosphate In Bacteriasupporting

confidence: 62%

Hydrolysis and synthesis of thiamin triphosphate in bacteria.

Nishimune

Hayashi

1987

Journal of Nutritional Science and Vitaminology, J Nutr Sci Vit

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“…Using this gradient, TPP, TMP and thiamine could be well separated (Figure 1A). The addition of 1.5% N,N-dimethylformamide (DMF) to buffer A, reported to result in better separation [29], improved neither the separation nor the peak shape of the three thiochromes in our hands. Moreover, the baseline increased during elution when DMF was present.…”

Section: Methodsmentioning

confidence: 68%

Presence of thiamine pyrophosphate in mammalian peroxisomes

et al. 2007

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BackgroundThiamine pyrophosphate (TPP) is a cofactor for 2-hydroxyacyl-CoA lyase 1 (HACL1), a peroxisomal enzyme essential for the α-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.ResultsBecause 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.ConclusionOur 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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“…The finding that vertebrate tissues (especially the liver) contain adenylated thiamine compounds may lead us to re‐examine and in some cases question the validity of earlier reports concerning the exact ThTP content of some tissues or the enzymatic mechanisms of ThTP synthesis. For example, several authors [19–22] have claimed that the rat liver had a ThTP content several times higher than the brain. From our data (see Fig.…”

Section: Discussionmentioning

confidence: 99%

Thiaminylated adenine nucleotides. Chemical synthesis, structural characterization and natural occurrence

et al. 2009

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Thiamine and its three phosphorylated derivatives (mono‐, di‐ and triphosphate) occur naturally in most cells. Recently, we reported the presence of a fourth thiamine derivative, adenosine thiamine triphosphate, produced in Escherichia coli in response to carbon starvation. Here, we show that the chemical synthesis of adenosine thiamine triphosphate leads to another new compound, adenosine thiamine diphosphate, as a side product. The structure of both compounds was confirmed by MS analysis and 1H‐, 13C‐ and 31P‐NMR, and some of their chemical properties were determined. Our results show an upfield shifting of the C‐2 proton of the thiazolium ring in adenosine thiamine derivatives compared with conventional thiamine phosphate derivatives. This modification of the electronic environment of the C‐2 proton might be explained by a through‐space interaction with the adenosine moiety, suggesting U‐shaped folding of adenosine thiamine derivatives. Such a structure in which the C‐2 proton is embedded in a closed conformation can be located using molecular modeling as an energy minimum. In E. coli, adenosine thiamine triphosphate may account for 15% of the total thiamine under energy stress. It is less abundant in eukaryotic organisms, but is consistently found in mammalian tissues and some cell lines. Using HPLC, we show for the first time that adenosine thiamine diphosphate may also occur in small amounts in E. coli and in vertebrate liver. The discovery of two natural thiamine adenine compounds further highlights the complexity and diversity of thiamine biochemistry, which is not restricted to the cofactor role of thiamine diphosphate.

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Reversed-phase high-performance liquid chromatographic analysis of thiamine phosphate esters at subpicomole levels

Cited by 39 publications

References 4 publications

Hydrolysis and synthesis of thiamin triphosphate in bacteria.

Hydrolysis and synthesis of thiamin triphosphate in bacteria.

Presence of thiamine pyrophosphate in mammalian peroxisomes

Thiaminylated adenine nucleotides. Chemical synthesis, structural characterization and natural occurrence

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