Properties of 130 kDa subunit of monkey aldehyde oxidase.
We previously demonstrated the existence of a minor 130 kDa subunit in the sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)/Western blot analysis of monkey liver cytosol and expressed monkey aldehyde oxidase (AO) in Escherichia coli. In contrast, the 130 kDa subunit was not observed in rat AO. In the current study, the properties of the 130 kDa subunit were investigated from the viewpoint of species differences in the presence of the subunit and AO activity. Monkey AO with His-tag at the N- and C-terminus were expressed, and were immunoanalyzed with anti-AO and anti-His-tag antisera. The results revealed that the minor 130 kDa subunit was produced by cleavage at the N-terminal side of the 150 kDa subunit. The cleavage point was shown to be located between 188Leu and 189Pro of 150 kDa AO subunit by the Edman degradation method. The two amino acids related to the cleavage are contained in the linkage between the 2Fe-2S and FAD domains in AO of human and monkey, but not in AO of rat and mouse. As a fact, the 130 kDa subunit was observed in AO of human and monkey, but not in AO of rat and mouse, suggesting the two amino acids might be one reason of a species difference in the formation of the 130 kDa subunit. However, the existence of the 130 kDa subunit is not associated with the species differences in AO activity, because the cleavage results in the loss of 2Fe-2S cluster domain essential for exertion of AO activity.
Asakawa
Tasuku
T
Department of Drug Metabolism and Pharmacokinetics, Tohoku Pharmaceutical University, 4-4-1 Komatsushima, Aoba-ku, Sendai 981-8558, Japan.
Itoh
Kunio
K
Adachi
Mayuko
M
Hoshino
Kouichi
K
Watanabe
Nobuaki
N
Tanaka
Yorihisa
Y
D042931:Aldehyde Oxidase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D015153:Blotting, Western; D003001:Cloning, Molecular; D003600:Cytosol / Q000201:enzymology; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics; D000882:Haplorhini; D006801:Humans; D008099:Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D011487:Protein Conformation; D021122:Protein Subunits; D013045:Species Specificity
D021122:Protein Subunits; D042931:Aldehyde Oxidase