The location of the ligand-binding site of carbohydrate-binding modules that have evolved from a common sequence is not conserved

被引:104
作者
Czjzek, M
Bolam, DN
Mosbah, A
Allouch, J
Fontes, CMGA
Ferreira, LMA
Bornet, O
Zamboni, V
Darbon, H
Smith, NL
Black, GW
Henrissat, B
Gilbert, HJ
机构
[1] CNRS, Lab Architecture & Fonct Macromol Biol, IBSM, F-13402 Marseille 20, France
[2] Univ Aix Marseille 1, F-13402 Marseille 20, France
[3] Univ Aix Marseille 2, F-13402 Marseille 20, France
[4] Univ Newcastle Upon Tyne, Dept Biol & Nutr Sci, Newcastle Upon Tyne NE1 7RU, Tyne & Wear, England
[5] Fac Med Vet, CIISA, P-1199 Lisbon, Portugal
[6] Northumbria Univ, Sch Appl & Mol Sci, Newcastle Upon Tyne NE1 8ST, Tyne & Wear, England
关键词
D O I
10.1074/jbc.M109142200
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Polysaccharide-degrading enzymes are generally modular proteins that contain non-catalytic carbohydrate-binding modules (CBMs), which potentiate the activity of the catalytic module. CBMs have been grouped into sequence-based families, and three-dimensional structural data are available for half of these families. Clostridium thermocellum xylanase 11A is a modular enzyme that contains a CBM from family 6 (CBM6), for which no structural data are available. We have determined the crystal structure of this module to a resolution of 2.1 Angstrom. The protein is a beta -sandwich that contains two potential ligand-binding clefts designated cleft A and B. The CBM interacts primarily with xylan, and NMR spectroscopy coupled with site-directed mutagenesis identified cleft A, containing Trp-92, Tyr-34, and Asn-120, as the ligand-binding site. The overall fold of CBM6 is similar to proteins in CBM families 4 and 22, although surprisingly the ligand-binding site in CBM4 and CBM22 is equivalent to cleft B in CBM6. These structural data define a superfamily of CBMs, comprising CBM4, CBM6, and CBM22, and demonstrate that, although CBMs have evolved from a relatively small number of ancestors, the structural elements involved in ligand recognition have been assembled at different locations on the ancestral scaffold.
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页码:48580 / 48587
页数:8
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