Substituting selenocysteine for active site cysteine 149 of phosphorylating glyceraldehyde 3-phosphate dehydrogenase reveals a peroxidase activity.

Abstract:

:Replacing the essential Cys-149 by a selenocysteine into the active site of phosphorylating glyceraldehyde 3-phosphate dehydrogenase (GAPDH) from Bacillus stearothermophilus leads to a selenoGAPDH that mimics a selenoperoxidase activity. Saturation kinetics were observed with cumenyl and tert-butyl hydroperoxides, with a better catalytic efficiency for the aromatic compound. The enzymatic mechanism fits a sequential model where the formation of a ternary complex between the holoselenoenzyme, the 3-carboxy 4-nitrobenzenethiol used as the reductant and the hydroperoxide precedes product release. The fact that the selenoGAPDH is NAD-saturated supports a binding of hydroperoxide and reductant in the substrate binding site. The catalytic efficiency is similar to selenosubtilisins but remains low compared to selenoglutathione peroxidase. This is discussed in relation to what is known from the X-ray crystal structures of selenoglutathione peroxidase and GAPDHs.

journal_name

FEBS Lett

journal_title

FEBS letters

authors

Boschi-Muller S,Muller S,Van Dorsselaer A,Böck A,Branlant G

doi

10.1016/s0014-5793(98)01377-5

subject

Has Abstract

pub_date

1998-11-20 00:00:00

pages

241-5

issue

3

eissn

0014-5793

issn

1873-3468

pii

S0014-5793(98)01377-5

journal_volume

439

pub_type

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