The O-glycosylated linker from the Trichoderma reesei Family 7 cellulase is a flexible, disordered protein.
Fungi and bacteria secrete glycoprotein cocktails to deconstruct cellulose. Cellulose-degrading enzymes (cellulases) are often modular, with catalytic domains for cellulose hydrolysis and carbohydrate-binding modules connected by linkers rich in serine and threonine with O-glycosylation. Few studies have probed the role that the linker and O-glycans play in catalysis. Since different expression and growth conditions produce different glycosylation patterns that affect enzyme activity, the structure-function relationships that glycosylation imparts to linkers are relevant for understanding cellulase mechanisms. Here, the linker of the Trichoderma reesei Family 7 cellobiohydrolase (Cel7A) is examined by simulation. Our results suggest that the Cel7A linker is an intrinsically disordered protein with and without glycosylation. Contrary to the predominant view, the O-glycosylation does not change the stiffness of the linker, as measured by the relative fluctuations in the end-to-end distance; rather, it provides a 16 Å extension, thus expanding the operating range of Cel7A. We explain observations from previous biochemical experiments in the light of results obtained here, and compare the Cel7A linker with linkers from other cellulases with sequence-based tools to predict disorder. This preliminary screen indicates that linkers from Family 7 enzymes from other genera and other cellulases within T. reesei may not be as disordered, warranting further study.
National Bioenergy Center, National Renewable Energy Laboratory, Golden, Colorado, USA Gregg.Beckham@nrel.gov
Gregg T
GT
Beckham
Yannick J
YJ
Bomble
James F
JF
Matthews
Courtney B
CB
Taylor
Michael G
MG
Resch
John M
JM
Yarbrough
Steve R
SR
Decker
Lintao
L
Bu
Xiongce
X
Zhao
Clare
C
McCabe
Jakob
J
Wohlert
Malin
M
Bergenstråhle
John W
JW
Brady
William S
WS
Adney
Michael E
ME
Himmel
Michael F
MF
Crowley
D000595:Amino Acid Sequence; D002480:Cellulase / Q000737:chemistry* / Q000378:metabolism*; D006031:Glycosylation; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D013816:Thermodynamics; D014242:Trichoderma / Q000201:enzymology*
D002480:Cellulase