Substrate specificity and subsite mobility in T. aurantiacus xylanase 10A.
The substrate specificity of Thermoascus aurantiacus xylanase 10A (TAX) has been investigated both biochemically and structurally. High resolution crystallographic analyses at 291 K and 100 K of TAX complexes with xylobiose show that the ligand is in its alpha anomeric conformation and provide a rationale for specificity on p-nitrophenyl glycosides at the -1 and -2 subsites. Trp 275, which is disordered in uncomplexed structures, is stabilised by its interaction with xylobiose. Two structural subsets in family 10 are identified, which differ by the presence or absence of a short helical stretch in the eighth betaalpha-loop of the TIM barrel, the loop bearing Trp 275. This structural difference is discussed in the context of Trp 275 mobility and xylanase function.
Lo Leggio
L
L
Centre for Crystallographic Studies, Chemical Institute, University of Copenhagen, Universitetsparken 5, DK-2100 Copenhagen, Denmark. leila@ccs.ki.ku.dk
Kalogiannis
S
S
Eckert
K
K
Teixeira
S C
SC
Bhat
M K
MK
Andrei
C
C
Pickersgill
R W
RW
Larsen
S
S
D001203:Ascomycota / Q000201:enzymology*; D020134:Catalytic Domain; D004187:Disaccharides / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D008958:Models, Molecular; D018716:Molecular Mimicry; D009038:Motion; D013379:Substrate Specificity; D014364:Tryptophan; D043325:Xylan Endo-1,3-beta-Xylosidase; D014995:Xylosidases / Q000378:metabolism*
D004187:Disaccharides; D014364:Tryptophan; D014995:Xylosidases; D043325:Xylan Endo-1,3-beta-Xylosidase; C004173:xylobiose; D005990:Glycerol