Native acridone synthases I and II from Ruta graveolens L. form homodimers

FEBS Letters

Springob, Karin³  Urbanke, Claus²  Lukačin, Richard³  Ernwein, Claudia³  Matern, Ulrich³  Schröder, Joachim¹  Schröder, Gudrun¹ 

[1] Biologisches Institut II, Albert-Ludwigs-Universität, Schänzlestrasse 1, D-79104 Freiburg, Germany;Medizinische Hochschule Hannover, Betriebseinheit Biophysikalisch-Biochemische Verfahren, Carl Neuberg Str. 1, 30623 Hannover, Germany;Institut für Pharmazeutische Biologie, Universität Marburg, Deutschhausstrasse 17A, D-35037 Marburg, Germany

关键词: Acridone synthase;    Polyketide synthase;    Subunit composition;    Acridone alkaloid biosynthesis;    Ruta graveolens L;    ACS;    acridone synthase;    CHS;    chalcone synthase;    ORF;    open reading frame;    IPTG;    isopropyl-β-d-thiogalactoside;    SDS-PAGE;    polyacrylamide gel electrophoresis in the presence of sodium dodecylsulfate;   

DOI  :  10.1016/S0014-5793(99)00355-5

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Acridone synthase II cDNA was cloned from irradiated cell suspension cultures of Ruta graveolens L. and expressed in Escherichia coli. The translated polypeptide of M _r 42 681 revealed a high degree of similarity to heterologous chalcone and stilbene synthases (70–75%), and the sequence was 94% identical to that of acridone synthase I cloned previously from elicited Ruta cells. Highly active recombinant acridone synthases I and II were purified to apparent homogeneity by a four-step purification protocol, and the affinities to N-methylanthraniloyl-CoA and malonyl-CoA were determined. The molecular mass of acridone synthase II was estimated from size exclusion chromatography on a Fractogel EMD BioSEC (S) column at about 45 kDa, as compared to a mass of 44±3 kDa found for the acridone synthase I on Superdex 75. Nevertheless, the sedimentation analysis by ultracentrifugation revealed molecular masses of 81±4 kDa for both acridone synthases. It is proposed, therefore, that the acridone synthases of Ruta graveolens are typical homodimeric plant polyketide synthases.

【 授权许可】

Unknown