The cDNA sequence and the deduced amino acid sequence of human transcobalamin II show homology with rat intrinsic factor and human transcobalamin I.
The cellular uptake of cobalamin (Cbl, vitamin B12) is mediated by transcobalamin II (TCII), a plasma protein that binds Cbl and is secreted by human umbilical vein endothelial (HUVE) cells. These cells synthesize and secrete TCII and, therefore, served as the source of the complementary DNA (cDNA) library from which the TCII cDNA was isolated. This full-length cDNA consists of 1866 nucleotides that code for a leader peptide of 18 amino acids, a secreted protein of 409 amino acids, a 5'-untranslated segment of 37 nucleotides, and a 3'-untranslated region of 548 nucleotides. A single 1.9-kilobase species of mRNA corresponding to the size of the cDNA was identified by Northern blot analysis of the RNA isolated from HUVE cells. TCII has 20% amino acid homology and greater than 50% nucleotide homology with human transcobalamin I (TCI) and with rat intrinsic factor (R-IF). TCII has no homology with the amino-terminal region of R-IF that has been reported to have significant primary as well as secondary structural homology with the nucleotide-binding domain of NAD-dependent oxidoreductases. The regions of homology that are common to all three proteins are located in seven domains of the amino acid sequence. One or more of these conserved domains is likely to be involved in Cbl binding, a function that is common to all three proteins. However, the difference in the affinity of TCII, TCI, and R-IF for Cbl and Cbl analogues indicates, a priori, that structural differences in the ligand-binding site of these proteins exist and these probably resulted from divergence of a common ancestral gene.
Platica
O
O
Department of Medicine, State University of New York Health Science Center, Brooklyn.
Janeczko
R
R
Quadros
E V
EV
Regec
A
A
Romain
R
R
Rothenberg
S P
SP
D000595:Amino Acid Sequence; D000818:Animals; D015152:Blotting, Northern; D004247:DNA / Q000235:genetics*; D006801:Humans; D007437:Intrinsic Factor / Q000235:genetics*; D008969:Molecular Sequence Data; D011061:Poly A / Q000032:analysis; D012313:RNA / Q000032:analysis; D051381:Rats; D015183:Restriction Mapping; D012689:Sequence Homology, Nucleic Acid; D014155:Transcobalamins / Q000235:genetics*
D014155:Transcobalamins; D011061:Poly A; D012313:RNA; D004247:DNA; D007437:Intrinsic Factor