cDNA cloning and characterization of human glyoxalase I isoforms from HT-1080 cells.
We have isolated two kinds of cDNA clones encoding glyoxalase I from a human fibrosarcoma HT-1080 cDNA library. One of them is identical to the glyoxalase I cDNA isolated by us, and the other encodes a protein in which alanine at position 111 of the reported sequence for glyoxalase I is replaced by glutamic acid. When the two cDNAs were co-translated in vitro, three bands representing two homodimers and one heterodimer appeared on native polyacrylamide gel electrophoresis, as observed for glyoxalase I purified from human erythrocytes or derived from a HT-1080 cell lysate. Escherichia coli cells carrying an expression vector of one of the novel glyoxalase I cDNAs showed glyoxalase I activity. These results reveal that two isoforms of human glyoxalase I showing different electrophoretic properties result from a change in one amino acid residue.
Kim
N S
NS
Kanagawa Academy of Science and Technology.
Sekine
S
S
Kiuchi
N
N
Kato
S
S
D015152:Blotting, Northern; D002460:Cell Line; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000378:metabolism; D005354:Fibrosarcoma; D015723:Gene Library; D006801:Humans; D007527:Isoenzymes / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D007791:Lactoylglutathione Lyase / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D046911:Macromolecular Substances; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D007527:Isoenzymes; D046911:Macromolecular Substances; D012333:RNA, Messenger; D007791:Lactoylglutathione Lyase