Structure of a Talaromyces pinophilus GH62 arabinofuranosidase in complex with AraDNJ at 1.25 Å resolution

Abstract

The three-dimensional structure of a fungal arabinofuranosidase from CAZY family GH62 has been solved at 1.25 Å resolution in complex with the bespoke arabinofuranosidase inhibitor AraDNJ, shedding light on the activity of this catalyst in the enzymatic deconstruction of arabinoxylans.

Structure of a GH51 α-L-arabinofuranosidase from Meripilus giganteus: conserved substrate recognition from bacteria to fungi

“…AraDNJ is a well known reversible -l-arabinofuranosidase inhibitor. Often thought to mimic the oxocarbeniumion transition state of -l-arabinofuranoside cleavage (Gloster et al, 2007), this inhibitor has recently been applied in the understanding of the mechanism of GH62 -l-arabinofuranosidases (Moroz et al, 2018), where it bound in a Michaelis complex-like 4 E conformation. Solving the structure of MgGH51 in the presence of AraDNJ gave a 1.70 Å resoljution structure with clear density for AraDNJ in the active site.…”

“…Chemical biology tools provide a more facile approach to the study of fungal enzymes, which require considerably more work to genetically engineer. For example, reversible inhibitors such as 1,4-dideoxy-1,4-imino-l-arabinitol (AraDNJ) have been applied to the study of GH62 -l-arabinofuranosidases, binding in a Michaelis complex-like 4 E conformation and facilitating the identification of the catalytic water molecule of an inverting -l-arabinofuranosidase (Moroz et al, 2018). Irreversible inhibitors, such as -larabinocyclophellitol aziridine (-l-AraAZI) and -l-arabinocyclophellitol cyclic sulfate (-l-AraCS), have recently been shown to label the active sites of both GH51 and GH54 -l-arabinofuranosidases, often mimicking the expected 2 E conformation of the glycosyl-enzyme intermediate (McGregor et al, 2020).…”

Carbohydrate structure hits the groove

“…In the remainder of the special issue we publish the structures of the highly glycosylated human leukocyte elastase in complex with an inhibitor (Hochscherf et al, 2018), of several enzymes that act on carbohydrates (Moroz et al, 2018;Urresti et al, 2018;Wu & Davies, 2018) and of a transcriptional regulator bound to a glycan effector (Mahounga et al, 2018).…”