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A novel 17 beta-hydroxysteroid dehydrogenase in the fungus Cochliobolus lnatus: new insights into the evolution of steroid-hormone signallingLanišnik-Rižner, Tea ...17beta-Hydroxysteroid dehydrogenase (17beta-HSD) from the filamentous fungus Cochliobolus lunatus (17beta-HSDcl) catalyses the reduction of steroids and ofseveral o- and p-quinones. After ... purification of the enzyme, its partial amino acid sequence was determined. A PCR fragment amplified with primers derived from peptide sequences was generated for screening the Coch. lunatus cDNA library. Three independent full-length cDNA clones were isolated and sequenced, revealing an 810-bp open reading frame encoding a 270-amino-acid protein. After expression in Escherichia coli and purification to homogeneity,the enzyme was found to be active towards androstenedione and menadione, and was able to form dimers of Mr 60000. The amino acid sequence ofthe novel 17beta-HSD demonstrated high homology with fungal carbonyl reductases, such as versicolorin reductase from Emericella nidulans (Aspergillus nidulans; VerA) and Asp. parasiticus (Ver1), polyhydroxynaphthalene reductase from Magnaporthe grisea, the product of the Brn 1 gene from Coch. heterostrophus and a reductase from Colletotrichum lagenarium, which are all members of the short-chain dehydrogenase/ reductase superfamily. 17beta-HSDcl is the first discovered fungal 17beta-hydroxysteroiddehydrogenase belonging to this familv. The primary structure of this enzyme may therefore help to elucidate the evolutionary history of steroid dehydrogenases.Vir: Biochemical journal. - ISSN 0264-6021 (Letn. 337, 1999, str. 425-431)Vrsta gradiva - članek, sestavni delLeto - 1999Jezik - angleškiCOBISS.SI-ID - 10122713
Avtor
Lanišnik-Rižner, Tea |
Moeller, Gabriele |
Thole, Hubert H. |
Žakelj-Mavrič, Marija |
Adamski, Jerzy, molekularna biologija
Teme
Ascomycetes |
Metabolism |
Enzymology |
Steroids |
17-hydroxysteroid dehydrogenases |
Biosynthesis |
Metabolism |
Polymerase chain reaction |
Vitamin K |
Metabolism |
Phylogeny |
Molecular weight |
Escherichia coli |
Genetics |
Metabolism |
Amino acid sequence |
Androstenedione |
Base sequence |
DNA, complementary |
Genetics |
Isolation and purification |
Electrophoresis, polyacrylamide gel
Vnos na polico
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Lanišnik-Rižner, Tea | 11699 |
Žakelj-Mavrič, Marija | 03082 |
Adamski, Jerzy, molekularna biologija | 55378 |
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