Seabream antiquitin: molecular cloning, tissue distribution, subcellular localization and functional expression.
Subsequent to our earlier report on the first purification of antiquitin protein from seabream liver and demonstration of its enzymatic activity [FEBS Letters 516 (2002) 183-186], we report herein the cloning of its full-length cDNA sequence. The open reading frame encodes a protein of 511 amino acids. Results of RT-PCR indicate that antiquitin is highly expressed in both the seabream liver and kidney. Transfection studies in cultured eukaryotic cells provided further evidence that it is a cytosolic protein. Bacterial expression of the enzyme was also performed. The purified recombinant protein was demonstrated to exhibit similar kinetic properties as the native enzyme.
Tang
Wai-Kwan
WK
Department of Biochemistry, The Chinese University of Hong Kong, Shatin, NT, Hong Kong, China.
Chan
Chi-Bun
CB
Cheng
Christopher H K
CH
Fong
Wing-Ping
WP
D000444:Aldehyde Dehydrogenase / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D029941:Fish Proteins / Q000032:analysis / Q000235:genetics* / Q000378:metabolism*; D007668:Kidney / Q000737:chemistry / Q000378:metabolism; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008928:Mitochondria / Q000737:chemistry; D008969:Molecular Sequence Data; D010802:Phylogeny; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D021541:Sea Bream / Q000235:genetics / Q000378:metabolism*; D014018:Tissue Distribution
D029941:Fish Proteins; D011994:Recombinant Proteins; D000444:Aldehyde Dehydrogenase