Molecular cloning and tissue distribution of rat sarcosine dehydrogenase

被引:35
作者
Bergeron, F
Otto, A
Blache, P
Day, R
Denoroy, L
Brandsch, R
Bataille, D [1 ]
机构
[1] CHU Arnaud de Villeneuve, INSERM U376, F-34295 Montpellier 05, France
[2] Univ Freiburg, Inst Biochem, Freiburg, Germany
[3] Inst Pharmacol Sherbrooke, Sherbrooke, PQ, Canada
[4] CNRS, Serv Cent Anal, F-69390 Vernaison, France
来源
EUROPEAN JOURNAL OF BIOCHEMISTRY | 1998年 / 257卷 / 03期
关键词
sarcosine dehydrogenase; dimethylglycine dehydrogenase; metabolism of one-carbon amino acids; tissue distribution;
D O I
10.1046/j.1432-1327.1998.2570556.x
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Sarcosine dehydrogenase (SarDH) is a mitochondrial flavoenzyme involved in the oxidative degradation of choline to glycine. The absence of SarDH activity in humans is genetically transmitted and is the cause of an amino acid metabolism disorder called sarcosinemia. Tryptic fragments of the purified enzyme from rat liver were subjected to Edman degradation and the sequences obtained were used to clone the cDNA encoding the full length protein. The deduced amino acid sequence of SarDH shares an overall similarity of 47% with dimethylglycine dehydrogenase (Me(2)GlyDH), another flavoenzyme involved in the mitochondrial choline catabolism with a similar FAD-binding domain. Covalent binding of FAD to SarDH was demonstrated by the observation of strong fluorescence at 530 nm under excitation at 450 nm of the enzyme immunoprecipitated under denaturing conditions from liver extracts. The localization of SarDH immunoreactivity in the mitochondrial matrix was confirmed by Western-blot analysis of purified mitochondrial fractions. Finally, the tissue distribution of SarDH was investigated by Northern-blot analysis of total RNA and Western-blot analysis of total protein from several rat tissues. A strong expression in the liver, but also in the lung, pancreas, kidney, thymus, and oviduct was observed. We therefore suggest that the enzymes of the choline catabolism pathway are important also for metabolism in nonhepatic tissues.
引用
收藏
页码:556 / 561
页数:6
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