Expression of alpha-amylase gene in rat liver: liver-specific amylase has a high affinity to glycogen.
The reactivity of rat liver alpha-amylases with maltotriose (G3), maltopentose (G5) and glycogen has been investigated. Liver amylases were found to be glycosylated and to have a molecular mass of 50 kDa by Western blotting using an anti-human salivary amylase antibody. The glycosylated liver amylases were found to be capable of G3- and G5-hydrolysis and of glucose formation, as demonstrated by thin-layer chromatography. When the amylase preparation was exposed to different concentrations of glycogen and run on a cellulose acetate membrane, the mobilities of rat liver amylases significantly decreased with tailing directly from the point of origin. In contrast, rat salivary amylases were not so much. These results indicate that rat liver amylases have a strong affinity to glycogen. We confirmed the expression of liver-specific amylases in rat liver by reverse transcriptional-polymerase chain reaction (RT-PCR); PCR products showed one band of an expected size of 474 bp using primers tested in the present study. A partial nucleotide sequence was then determined. When compared with the gene of mouse liver amylase, the substitution of 26 bases out of 434 bases was elucidated. The present data demonstrate the presence of liver-specific amylases in rats.
Koyama
I
I
Department of Medical Technology, Junior College, Saitama Medical School, Saitama, Japan. koyamaw@saitama-med.ac.jp
Komine
S
S
Hokari
S
S
Yakushijin
M
M
Matsunaga
T
T
Komoda
T
T
D000818:Animals; D001483:Base Sequence; D002241:Carbohydrates; D015870:Gene Expression; D006003:Glycogen / Q000378:metabolism*; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012333:RNA, Messenger; D051381:Rats; D017208:Rats, Wistar; D013379:Substrate Specificity; D000516:alpha-Amylases / Q000235:genetics* / Q000378:metabolism
D002241:Carbohydrates; D012333:RNA, Messenger; D006003:Glycogen; D000516:alpha-Amylases