The primary structure of a novel riboflavin-binding protein of emu (Dromaius novaehollandiae).
Emu riboflavin-binding protein (RBP) was purified from egg white and yolk, and its N-terminal amino acid sequence was determined. The molecular mass of emu RBP was estimated at approximately 48 and 45 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, i.e., 10 kDa larger than chicken RBP. The molecular mass of deglycosylated RBPs indicated that the content of oligosaccharide chain in emu RBP was approximately 3 times greater than that in chicken RBP. The gene encoding the RBP precursor was cloned from emu oviduct cDNA by PCR and found also in the liver and ovary cDNAs as well as oviduct cDNA. The complete cDNA consisted of an open reading frame of 714 bp encoding a protein of 238 amino acids. The amino acid sequence deduced from the cDNA sequence revealed that many essential structural features were conserved in emu RBP including 18 cysteine residues, 2 N-glycosylation sites, a clustered phosphorylation region, and riboflavin-binding sites. Two additional potential N-glycosylation sites were found in the amino acid sequences of RBPs from the emu and other sources such as the turtle and frog, which might in part account for the greater content of oligosaccharide chain of emu RBP as compared to chicken RBP.
Maehashi
Kenji
K
Department of Fermentation Science, Faculty of Applied Bio-Science, Tokyo University of Agriculture, Japan. maehashi@nodai.ac.jp
Matano
Mami
M
Uchino
Masataka
M
Yamamoto
Yasushi
Y
Takano
Katsumi
K
Watanabe
Toshihiro
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D020312:Dromaiidae / Q000235:genetics*; D004527:Egg Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D026901:Membrane Transport Proteins / Q000737:chemistry / Q000145:classification / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D017386:Sequence Homology, Amino Acid
D018076:DNA, Complementary; D004527:Egg Proteins; D026901:Membrane Transport Proteins; C019079:riboflavin-binding protein