Molecular cloning of cDNAs of human liver and placenta NADH-cytochrome b5 reductase.
A cDNA coding for human liver NADH-cytochrome b5 reductase (cytochrome b5 reductase, EC 1.6.2.2) was cloned from a human liver cDNA library constructed in phage lambda gt11. The library was screened by using an affinity-purified rabbit antibody against NADH-cytochrome b5 reductase of human erythrocytes. A cDNA about 1.3 kilobase pairs long was isolated. By using the cDNA as a probe, another cDNA (pb5R141) of 1817 base pairs was isolated that hybridized with a synthetic oligonucleotide encoding Pro-Asp-Ile-Lys-Tyr-Pro, derived from the amino acid sequence at the amino-terminal region of the enzyme from human erythrocytes. Furthermore, by using the pb5R141 as a probe, cDNA clones having more 5' sequence were isolated from a human placenta cDNA library. The amino acid sequences deduced from the nucleotide sequences of these cDNA clones overlapped each other and consisted of a sequence that completely coincides with that of human erythrocytes and a sequence of 19 amino acid residues extended at the amino-terminal side. The latter sequence closely resembles that of the membrane-binding domain of steer liver microsomal enzyme.
D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D003579:Cytochrome Reductases / Q000235:genetics*; D042966:Cytochrome-B(5) Reductase; D004247:DNA / Q000378:metabolism*; D004262:DNA Restriction Enzymes; D005260:Female; D006801:Humans; D008099:Liver / Q000201:enzymology*; D009693:Nucleic Acid Hybridization; D010920:Placenta / Q000201:enzymology*
D004247:DNA; D003579:Cytochrome Reductases; D042966:Cytochrome-B(5) Reductase; D004262:DNA Restriction Enzymes