The giant extracellular hemoglobin from the polychaete Neanthes diversicolor. The cDNA-derived amino acid sequence of linker chain L2 and the exon/intron boundary conserved in linker genes.
The 4000 kDa extracellular hemoglobin from the polychaete Neanthes diversicolor consists of three types of subunits; three 15 kDa monomers (chains M1, M2 and M3), a 45 kDa disulfide-bonded trimer of chains T1, T2 and T3, and two 50-55 kDa disulfide-bonded homodimeric linkers (chains L1 and L2). The latter linker subunits are essential for the assembly of the other heme-containing subunits, monomers and a trimer. The cDNA encoding the linker chain L2 was amplified by polymerase chain reaction (PCR), and the cDNA-derived amino acid sequence of 235 residues has been determined. The sequence showed 22-75% identity with other linker chains. All of the linker sequences examined so far have a highly conserved cysteine-rich segment at positions 89-130: Xaa3-Cys-Xaa6-Cys-Xaa6-Cys-Xaa6-Cys-Asp-Gly-X aa2-Asp-Cys-Xaa4-Asp-Glu-Xaa4-Cys, and the motif corresponds exactly to the cysteine-rich repeats of the ligand-binding domains of vertebrate low-density lipoprotein (LDL) receptors (Suzuki, T. and Riggs, A.F. (1993) J. Biol. Chem. 268, 13548-13555). A 287 bp intron interrupts the coding sequence of Neanthes L2 gene just at the N-terminal boundary of this motif, and the position of the splice junction was exactly conserved in Neanthes and Lumbricus linker genes. This suggests that the intron has been conserved for at least 450 million years in annelid linker genes. The evolutionary origin of the remaining parts of linker chains is unclear, but it is noteworthy that the topology of the two intrachain disulfide bridges in the C-terminal segment of linker chains is homologous with that of the carbohydrate-recognition domain of animal C-type lectin.
Suzuki
T
T
Department of Biology, Faculty of Science, Kochi University, Japan.
Ohta
T
T
Yuasa
H J
HJ
Takagi
T
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001798:Blood Proteins / Q000737:chemistry / Q000235:genetics*; D018076:DNA, Complementary / Q000096:biosynthesis; D006454:Hemoglobins / Q000235:genetics / Q000302:isolation & purification*; D008969:Molecular Sequence Data; D011077:Polychaeta / Q000378:metabolism*; D012333:RNA, Messenger / Q000302:isolation & purification; D016415:Sequence Alignment
D001798:Blood Proteins; D018076:DNA, Complementary; D006454:Hemoglobins; D012333:RNA, Messenger; C086641:hemoglobin linker chain L2, Neanthes diversicolor