PuSH - Publikationsserver des Helmholtz Zentrums München

Export: TXT Text RIS Endnote (RIS) BIB BibTeX
Kristan, K.* ; Adamski, J. ; Lanisnik Rizner, T.* ; Stojan, J.*

His164 regulates accessibility to the active site in fungal 17β-hydroxysteroid dehydrogenase.

Biochimie 89, 63-71 (2007)
DOI PMC
Open Access Green möglich sobald Postprint bei der ZB eingereicht worden ist.
17ß-Hydroxysteroid dehydrogenase from the fungus Cochliobolus lunatus (17ß-HSDcl) is an NADPH-dependent member of the short-chain dehydrogenase/ reductase superfamily. To study the catalytic properties of this enzyme, we prepared several specific mutations of 17ß-HSDcl (Tyr167Phe, His164Trp/Gly, Tyr212Ala). Wild-type 17ß-HSDcl and the 17ß-HSDcl mutants were evaluated by chromatographic, kinetic and thermodynamic means. The Tyr167Phe mutation resulted in a complete loss of enzyme activity, while substitution of His164 with Trp and Gly both resulted in higher specificity number (V/K) for the steroid substrates, which are mainly a consequence of easier accessibility of steroid substrates to the active-site hollow under optimized conditions. The Tyr212Ala mutant showed increased activity in the oxidative direction, which appears to be a consequence of increased NADPH dissociation. The kinetic characterizations and thermodynamic analyses also suggest that His164 and Tyr212 in 17ß-HSDcl have a role in the opening and closing of the active site of this enzyme and in the discrimination between oxidized and reduced coenzyme.
Impact Factor
Scopus SNIP
Web of Science
Times Cited
Scopus
Cited By
Altmetric
3.237
0.000
8
8
Tags
Anmerkungen
Besondere Publikation
Auf Hompepage verbergern

Zusatzinfos bearbeiten
Eigene Tags bearbeiten
Privat
Eigene Anmerkung bearbeiten
Privat
Auf Publikationslisten für
Homepage nicht anzeigen
Als besondere Publikation
markieren
Publikationstyp Artikel: Journalartikel
Dokumenttyp Wissenschaftlicher Artikel
Schlagwörter 17ß-Hydroxysteroid dehydrogenase; Short-chain dehydrogenase/reductase; Site-directed mutagenesis; Kinetic characterization; Thermodynamic analysis
Sprache
Veröffentlichungsjahr 2007
HGF-Berichtsjahr 2006
ISSN (print) / ISBN 0300-9084
e-ISSN 1638-6183
Zeitschrift Biochimie
Quellenangaben Band: 89, Heft: 1, Seiten: 63-71 Artikelnummer: , Supplement: ,
Verlag Elsevier
Begutachtungsstatus Peer reviewed
Institut(e) Molekulare Endokrinologie und Metabolismus (MEM)
POF Topic(s) 30201 - Metabolic Health
Forschungsfeld(er) Genetics and Epidemiology
PSP-Element(e) G-505600-001
PubMed ID 17034927
DOI 10.1016/j.biochi.2006.09.004
WOS ID 000243630100006
Scopus ID 33845621427
Erfassungsdatum 2006-09-26
[➜Korrektur melden]