Production of homo- and hetero-dimeric isozymes from two aldehyde oxidase genes of Arabidopsis thaliana

被引:41
作者
Akaba, S
Seo, M
Dohmae, N
Takio, K
Sekimoto, H
Kamiya, Y
Furuya, N
Komano, T
Koshiba, T [1 ]
机构
[1] Tokyo Metropolitan Univ, Dept Biol, Tokyo 1920397, Japan
[2] RIKEN, Inst Phys & Chem Res, Div Biomol Characterizat, Wako, Saitama 3510198, Japan
[3] Univ Tokyo, Grad Sch Arts & Sci, Dept Life Sci, Tokyo 1538902, Japan
[4] RIKEN, Inst Phys & Chem Res, Frontier Res Program, Wako, Saitama 3510198, Japan
关键词
aldehyde oxidase; Arabidopsis thaliana; indole-3-acetic acid (IAA); isozyme; molybdenum cofactor;
D O I
10.1093/oxfordjournals.jbchem.a022463
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Polyclonal antibodies were raised against synthetic peptides or recombinant polypeptides encoded by Arabidopsis atAO-1 and atAO-2 a cDNAs, which have sequences similar to maize and animal aldehyde oxidase (AO) cDNAs. Anti-atAO-1 antibodies recognized AO alpha and AO beta among the three isoforms, AO alpha, AO beta, and AO gamma, detected in Arabidopsis seedlings after native PAGE, while anti-atAO-2 antibodies reacted with AO beta and AO gamma. 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 AOa, AO beta, and AO gamma were estimated as approximately 290-300 kDa. These results suggest that AO alpha and AO gamma are homodimers consisting of atAO-1 and atAO-2 products, respectively, and that AO beta is a heterodimer of the atAO-1 and atAO-2 products.
引用
收藏
页码:395 / 401
页数:7
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