Aminobutyric Acid Transporter 2 Mediates the Hepatic Uptake of Guanidinoacetate, the Creatine Biosynthetic Precursor, in Rats

Guanidinoacetic acid (GAA) is the biosynthetic precursor of creatine which is involved in storage and transmission of phosphate-bound energy. Hepatocytes readily convert GAA to creatine, raising the possibility that the active uptake of GAA by hepatocytes is a regulatory factor. The purpose of this...

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Bibliographic Details
Published in:PloS one 2012-02, Vol.7 (2), p.e32557
Main Authors: Tachikawa, Masanori, Ikeda, Saori, Fujinawa, Jun, Hirose, Shirou, Akanuma, Shin-ichi, Hosoya, Ken-ichi
Format: Article
Language:English
Subjects:
Creatine
GABA
Liver
Phosphates
Physiological aspects
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Summary:Guanidinoacetic acid (GAA) is the biosynthetic precursor of creatine which is involved in storage and transmission of phosphate-bound energy. Hepatocytes readily convert GAA to creatine, raising the possibility that the active uptake of GAA by hepatocytes is a regulatory factor. The purpose of this study is to investigate and identify the transporter responsible for GAA uptake by hepatocytes. The characteristics of [.sup.14 C]GAA uptake by hepatocytes were elucidated using the in vivo liver uptake method, freshly isolated rat hepatocytes, an expression system of Xenopus laevis oocytes, gene knockdown, and an immunohistochemical technique. In vivo injection of [.sup.14 C]GAA into the rat femoral vein and portal vein results in the rapid uptake of [.sup.14 C]GAA by the liver. The uptake was markedly inhibited by [gamma]-aminobutyric acid (GABA) and nipecotinic acid, an inhibitor of GABA transporters (GATs). The characteristics of Na.sup.+ - and Cl.sup.- -dependent [.sup.14 C]GAA uptake by freshly isolated rat hepatocytes were consistent with those of GAT2. The Km value of the GAA uptake (134 [micro]M) was close to that of GAT2-mediated GAA transport (78.9 [micro]M). GABA caused a marked inhibition with an IC.sub.50 value of 8.81 [micro]M. The [.sup.14 C]GAA uptake exhibited a significant reduction corresponding to the reduction in GAT2 protein expression. GAT2 was localized on the sinusoidal membrane of the hepatocytes predominantly in the periportal region. This distribution pattern was consistent with that of the creatine biosynthetic enzyme, S-adenosylmethionine:guanidinoacetate N-methyltransferase. GAT2 makes a major contribution to the sinusoidal GAA uptake by periportal hepatocytes, thus regulating creatine biosynthesis in the liver.
ISSN:1932-6203
1932-6203
DOI:10.1371/journal.pone.0032557