Molecular characterization and functional expression of squid retinal-binding protein. A novel species of hydrophobic ligand-binding protein.
The primary structure of squid retinal-binding protein (RALBP) was determined by cDNA and protein sequencing. Squid RALBP contains 342 amino acid residues in a single N-terminal-blocked chain with a molecular weight of 39,111. The N alpha-blocking group was identified as an acetyl moiety by mass spectrometry. The amino acid sequence revealed that the protein is highly hydrophilic and acidic, but it has several hydrophobic regions that are located mainly in the middle part of the polypeptide chain. It is also predicted that these hydrophobic regions form beta-sheet structures. The primary structure of RALBP is, however, quite distinct from those of other retinoid-binding proteins, showing that squid RALBP is a novel hydrophobic ligand-binding protein that functions in intracellular retinoid transport. Using the cloned cDNA, squid RALBP was expressed in vitro. By carrying out the translation at 20 degrees C in reticulocyte lysates, the protein having retinol binding activity was produced.
Ozaki
K
K
Department of Biology, Faculty of Science, Osaka University, Japan.
Terakita
A
A
Ozaki
M
M
Hara
R
R
Hara
T
T
Hara-Nishimura
I
I
Mori
H
H
Nishimura
M
M
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry* / Q000235:genetics; D003001:Cloning, Molecular; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D018076:DNA, Complementary / Q000378:metabolism; D049832:Decapodiformes / Q000378:metabolism*; D005123:Eye / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D011401:Promoter Regions, Genetic; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D015183:Restriction Mapping; D012172:Retinaldehyde / Q000378:metabolism
C020932:11-cis-retinal-binding protein; D002352:Carrier Proteins; D018076:DNA, Complementary; D010446:Peptide Fragments; D011994:Recombinant Proteins; D004247:DNA; D012172:Retinaldehyde