Structural enzymology studies with the substrate 3S‐hydroxybutanoyl‐CoA: bifunctional MFE1 is a less efficient dehydrogenase than monofunctional HAD DOI Creative Commons

Shruthi Sridhar,

Tiila‐Riikka Kiema, Werner Schmitz

et al.

FEBS Open Bio, Journal Year: 2024, Volume and Issue: 14(4), P. 655 - 674

Published: March 8, 2024

Multifunctional enzyme, type‐1 (MFE1) catalyzes the second and third step of β‐oxidation cycle, being, respectively, 2 E ‐enoyl‐CoA hydratase (ECH) reaction (N‐terminal part, crotonase fold) NAD + ‐dependent, 3 S ‐hydroxyacyl‐CoA dehydrogenase (HAD) (C‐terminal HAD fold). Structural enzymological properties rat MFE1 (RnMFE1) as well two its variants, namely E123A variant (a glutamate ECH active site is mutated into alanine) BCDE (without domain A part), were studied, using substrate ‐hydroxybutanoyl‐CoA. Protein crystallographic binding studies show hydrogen bond interactions ‐hydroxybutanoyl‐CoA 3‐keto, oxidized form, acetoacetyl‐CoA, with catalytic glutamates in site. Pre‐steady state experiments NADH that k on off rate constants monomeric RnMFE1 homologous human, dimeric (HsHAD) for are very similar, being same those observed variants. However, steady pre‐steady kinetic data concerning HAD‐catalyzed dehydrogenation that, cat chem conversion acetoacetyl‐CoA by (and variants) about 10 fold lower when catalyzed HsHAD. The dynamical dehydrogenases known to be important their efficiency, it discussed greater complexity correlates observation a slower than

Language: Английский

Polymorphic Structure Determination of the Macrocyclic Drug Paritaprevir by MicroED DOI Creative Commons
Guojun Bu, Emma Danelius, Lianne H. E. Wieske

et al.

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2023, Volume and Issue: unknown

Published: Sept. 10, 2023

Paritaprevir is an orally bioavailable, macrocyclic drug used for treating chronic Hepatitis C virus infection. Its structures had been elusive to the public until recently when one of crystal forms was solved by MicroED. In this work, we report MicroED two distinct polymorphic paritaprevir from same experiment. The different polymorphs show conformational changes in core, as well cyclopropylsulfonamide and methylpyrazinamide substituents. Molecular docking shows that conformations fits into active site pocket NS3/4A serine protease target, can interact with catalytic triad via hydrophobic interactions hydrogen bonds. These results provide further insight optimization binding acylsulfonamide inhibitors protease. addition, also demonstrate opportunity deriving macrocycle experiments using

Language: Английский

Citations

2
Structural enzymology studies with the substrate 3S‐hydroxybutanoyl‐CoA: bifunctional MFE1 is a less efficient dehydrogenase than monofunctional HAD DOI Creative Commons

Shruthi Sridhar,

Tiila‐Riikka Kiema, Werner Schmitz

et al.

FEBS Open Bio, Journal Year: 2024, Volume and Issue: 14(4), P. 655 - 674

Published: March 8, 2024

Multifunctional enzyme, type‐1 (MFE1) catalyzes the second and third step of β‐oxidation cycle, being, respectively, 2 E ‐enoyl‐CoA hydratase (ECH) reaction (N‐terminal part, crotonase fold) NAD + ‐dependent, 3 S ‐hydroxyacyl‐CoA dehydrogenase (HAD) (C‐terminal HAD fold). Structural enzymological properties rat MFE1 (RnMFE1) as well two its variants, namely E123A variant (a glutamate ECH active site is mutated into alanine) BCDE (without domain A part), were studied, using substrate ‐hydroxybutanoyl‐CoA. Protein crystallographic binding studies show hydrogen bond interactions ‐hydroxybutanoyl‐CoA 3‐keto, oxidized form, acetoacetyl‐CoA, with catalytic glutamates in site. Pre‐steady state experiments NADH that k on off rate constants monomeric RnMFE1 homologous human, dimeric (HsHAD) for are very similar, being same those observed variants. However, steady pre‐steady kinetic data concerning HAD‐catalyzed dehydrogenation that, cat chem conversion acetoacetyl‐CoA by (and variants) about 10 fold lower when catalyzed HsHAD. The dynamical dehydrogenases known to be important their efficiency, it discussed greater complexity correlates observation a slower than

Language: Английский

Citations

0