Transglycosylases strengthen the fungal cell wall by forming a rigid network of crosslinks. Here, Fang et al. show that the five Crh transglycosylases of Aspergillus fumigatus are dispensable for cell wall integrity in vitro, and solve the crystal structure of Crh5 in complex with chitooligosaccharides

A Gastebois

A Maddi

A Perrakis

A Pierleoni

A Vagin

AB Boraston

C d’Enfert

CS Rye

E Cabib

ECJ Arroyo

F Gomez-Esquer

G Pardini

GC Cerqueira

GN Murshudov

I Mouyna

J Arroyo

JM Rodriguez-Pena

JP Latge

LP Erwig

M Hahn

M Kopecka

M Mazan

ME Silva Ferreira da

N Blanco

NH Georgopapadakou

OJ Gaiser

P Emsley

P Jackson

P Johansson

S Armitt

SM Bowman

T Keitel

T Zelante

TG Battye

TN Petersen

V Aimanianda

WL DeLano

Fungal cell wall synthesis is achieved by a balance of glycosyltransferase, hydrolase and transglycosylase activities. Transglycosylases strengthen the cell wall by forming a rigid network of crosslinks through mechanisms that remain to be explored. Here we study the function of the Aspergillus fumigatus family of five Crh transglycosylases. Although crh genes are dispensable for cell viability, simultaneous deletion of all genes renders cells sensitive to cell wall interfering compounds. In vitro biochemical assays and localisation studies demonstrate that this family of enzymes functions redundantly as transglycosylases for both chitin-glucan and chitin-chitin cell wall crosslinks. To understand the molecular basis of this acceptor promiscuity, we solved the crystal structure of A. fumigatus Crh5 (AfCrh5) in complex with a chitooligosaccharide at the resolution of 2.8 Å, revealing an extensive elongated binding cleft for the donor (−4 to −1) substrate and a short acceptor (+1 to +2) binding site. Together with mutagenesis, the structure suggests a “hydrolysis product assisted” molecular mechanism favouring transglycosylation over hydrolysis.</p

Fang, Wenxia

Sanz, Ana Belén

Galan Bartual, Sergio

Wang, Bin

Ferenbach, Andrew

Farkaš, Vladimír

Hurtado-Guerrero, Ramón

Arroyo, Javier

van Aalten, Daan

University of Dundee Online Publications

English

Mechanisms of redundancy and specificity of the Aspergillus fumigatus Crh transglycosylases

Discovery Research Portal

Fungal cell wall synthesis is achieved by a balance of glycosyltransferase, hydrolase and transglycosylase activities. Transglycosylases strengthen the cell wall by forming a rigid network of crosslinks through mechanisms that remain to be explored. Here we study the function of the Aspergillus fumigatus family of five Crh transglycosylases. Although crh genes are dispensable for cell viability, simultaneous deletion of all genes renders cells sensitive to cell wall interfering compounds. In vitro biochemical assays and localisation studies demonstrate that this family of enzymes functions redundantly as transglycosylases for both chitin-glucan and chitin-chitin cell wall crosslinks. To understand the molecular basis of this acceptor promiscuity, we solved the crystal structure of A. fumigatus Crh5 (AfCrh5) in complex with a chitooligosaccharide at the resolution of 2.8 Å, revealing an extensive elongated binding cleft for the donor (-4 to -1) substrate and a short acceptor (+1 to +2) binding site. Together with mutagenesis, the structure suggests a "hydrolysis product assisted" molecular mechanism favouring transglycosylation over hydrolysis.Ministerio de Economía (MINECO)Comunidad de MadridDepto. de Microbiología y ParasitologíaFac. de FarmaciaTRUEpu

Sanz Santamaría, Ana Belén

Bartual, Sergio Galan

Ferenbach, Andrew T.

Hurtado Guerrero, Ramón

Van Aalten, Daan M. F.

Docta Complutense

Nature Communications

Mechanisms of redundancy and specificity of the Aspergillus fumigatus Crh transglycosylases.

Fungal cell wall synthesis is achieved by a balance of glycosyltransferase, hydrolase and transglycosylase activities. Transglycosylases strengthen the cell wall by forming a rigid network of crosslinks through mechanisms that remain to be explored. Here we study the function of the Aspergillus fumigatus family of five Crh transglycosylases. Although crh genes are dispensable for cell viability, simultaneous deletion of all genes renders cells sensitive to cell wall interfering compounds. In vitro biochemical assays and localisation studies demonstrate that this family of enzymes functions redundantly as transglycosylases for both chitin-glucan and chitin-chitin cell wall crosslinks. To understand the molecular basis of this acceptor promiscuity, we solved the crystal structure of A. fumigatus Crh5 (AfCrh5) in complex with a chitooligosaccharide at the resolution of 2.8 Å, revealing an extensive elongated binding cleft for the donor (-4 to -1) substrate and a short acceptor (+1 to +2) binding site. Together with mutagenesis, the structure suggests a “hydrolysis product assisted” molecular mechanism favouring transglycosylation over hydrolysis

Fang, W.

Sanz, A.B.

Bartual, S.G.

Wang, B.

Ferenbach, A.T.

Farkaš, V.

Hurtado-Guerrero, R.

Arroyo, J.

van Aalten, D.M.F.

Repositorio Universidad de Zaragoza

Crossref

http://link.springer.com/article/10.1038/s41467-019-09674-0

Mechanisms of redundancy and specificity of the Aspergillus fumigatus Crh transglycosylases

Abstract

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University of Dundee Online Publications

Discovery Research Portal

Docta Complutense

Discovery Research Portal

Repositorio Universidad de Zaragoza

Crossref