3-Keto-5 alpha -steroid Delta super(1)-dehydrogenase from Rhodococcus erythropolis SQ1 and its orthologue in Mycobacterium tuberculosis H37Rv are highly specific enzymes that function in cholesterol catabolism

The Rhodococcus erythropolis SQ1 kstD3 gene was cloned, heterologously expressed and biochemically characterized as a KSTD3 (3-keto- 5 alpha -steroid Delta super(1)-dehydrogenase). Upstream of kstD3, an ORF (open reading frame) with similarity to Delta super(4) KSTD (3-keto-5 alpha -steroid Delta su...

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Veröffentlicht in:Biochemical journal 2008-01, Vol.410 (1), p.339-346
Hauptverfasser: Knol, Jan, Bodewits, Karin, Hessels, Gerda I, Dijkhuizen, Lubbert, Van Der Geize, Robert
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Sprache:eng
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Zusammenfassung:The Rhodococcus erythropolis SQ1 kstD3 gene was cloned, heterologously expressed and biochemically characterized as a KSTD3 (3-keto- 5 alpha -steroid Delta super(1)-dehydrogenase). Upstream of kstD3, an ORF (open reading frame) with similarity to Delta super(4) KSTD (3-keto-5 alpha -steroid Delta super(4)- dehydrogenase) was found, tentatively designated kst4D. Biochemical analysis revealed that the Delta super(1) KSTD3 has a clear preference for 3- ketosteroids with a saturated A-ring, displaying highest activity on 5 alpha -AD (5 alpha -androstane-3,17-dione) and 5 alpha -T (5 alpha -testosterone; also known as 17 beta - hydroxy-5 alpha -androstane-3-one). The KSTD1 and KSTD2 enzymes, on the other hand, clearly prefer (9 alpha -hydroxy-)4-androstene-3,17-dione as substrates. Phylogenetic analysis of known and putative KSTD amino acid sequences showed that the R. erythropolis KSTD proteins cluster into four distinct groups. Interestingly, Delta super(1) KSTD3 from R. erythropolis SQ1 clustered with Rv3537, the only Delta super(1) KSTD present in Mycobacterium tuberculosis H37Rv, a protein involved in cholesterol catabolism and pathogenicity. The substrate range of heterologously expressed Rv3537 enzyme was nearly identical with that of Delta super(1) KSTD3, indicating that these are orthologous enzymes. The results imply that 5 alpha -AD and 5 alpha -T are newly identified intermediates in the cholesterol catabolic pathway, and important steroids with respect to pathogenicity.
ISSN:0264-6021
1470-8728