Production of homo- and hetero-dimeric isozymes from two aldehyde oxidase genes of Arabidopsis thaliana.
Polyclonal antibodies were raised against synthetic peptides or recombinant polypeptides encoded by Arabidopsis atAO-1 and atAO-2 cDNAs, which have sequences similar to maize and animal aldehyde oxidase (AO) cDNAs. Anti-atAO-1 antibodies recognized AOalpha and AObeta among the three isoforms, AOalpha, AObeta, and AOgamma, detected in Arabidopsis seedlings after native PAGE, while anti-atAO-2 antibodies reacted with AObeta and AOgamma. The polypeptide specifically recognized by each antibody was collected as the Protein-A/IgG/antigen complex. The 150- and 145-kDa polypeptides were purified by SDS-PAGE and digested with Achromobacter Protease I. From the amino acid sequences and molecular masses of the derivative peptides, it was revealed that the 150- and 145-kDa polypeptides were the products of atAO-1 and atAO-2, respectively. Molecular masses of the native forms of AOalpha, AObeta, and AOgamma were estimated as approximately 290-300 kDa. These results suggest that AOalpha and AOgamma are homodimers consisting of atAO-1 and atAO-2 products, respectively, and that AObeta is a heterodimer of the atAO-1 and atAO-2 products.
Akaba
S
S
Department of Biology, Tokyo Metropolitan University, Hachioji, Tokyo, 192-0397, Japan.
Seo
M
M
Dohmae
N
N
Takio
K
K
Sekimoto
H
H
Kamiya
Y
Y
Furuya
N
N
Komano
T
T
Koshiba
T
T
D042931:Aldehyde Oxidase; D000445:Aldehyde Oxidoreductases / Q000737:chemistry / Q000235:genetics*; D000595:Amino Acid Sequence; D000906:Antibodies / Q000378:metabolism; D017360:Arabidopsis / Q000201:enzymology* / Q000235:genetics*; D003067:Coenzymes; D017343:Genes, Plant; D007210:Indoleacetic Acids / Q000378:metabolism; D007527:Isoenzymes / Q000096:biosynthesis* / Q000737:chemistry; D008667:Metalloproteins / Q000378:metabolism; D008954:Models, Biological; D008957:Models, Genetic; D008969:Molecular Sequence Data; D000090006:Molybdenum Cofactors; D010455:Peptides / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics; D011233:Precipitin Tests; D011621:Pteridines / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
D042931:Aldehyde Oxidase; D000445:Aldehyde Oxidoreductases; D000906:Antibodies; D003067:Coenzymes; D007210:Indoleacetic Acids; D007527:Isoenzymes; D008667:Metalloproteins; D000090006:Molybdenum Cofactors; D010455:Peptides; D010940:Plant Proteins; D011621:Pteridines; D011994:Recombinant Proteins; C022261:molybdopterin