The structural basis for catalysis and specificity of the Pseudomonas cellulosa alpha-glucuronidase, GlcA67A

D  Nurizzo; T  Nagy; H J  Gilbert; G J  Davies

The structural basis for catalysis and specificity of the Pseudomonas cellulosa alpha-glucuronidase, GlcA67A

D Nurizzo, T Nagy, H J Gilbert, G J Davies

Chemistry

Research output: Contribution to journal › Article › peer-review

Abstract

alpha-glucuronidases, components of an ensemble of enzymes central to the recycling of photosynthetic biomass, remove the alpha-1,2 linked 4-O-methyl glucuronic acid from xylans. The structure of the alpha-glucuronidase, GlcA67A, from Pseudomonas cellulosa reveals three domains, the central of which is a (beta/alpha)(B) barrel housing the catalytic apparatus. Complexes of the enzyme with the individual reaction products, either xylobiose or glucuronic acid, and the ternary complex of both glucuronic acid and xylotriose reveal a "blind" pocket which selects for short decorated xylooligosaccharides substituted with the uronic acid at their nonreducing end, consistent with kinetic data. The catalytic center reveals a constellation of carboxylates; Glu292 is poised to provide protonic assistance to leaving group departure with Glu393 and Asp365 both appropriately positioned to provide base-catalyzed assistance for inverting nucleophilic attack by water.

Original language	English
Pages (from-to)	547-556
Number of pages	10
Journal	Structure
Volume	10
Issue number	4
Publication status	Published - Apr 2002

Keywords

glucuronidase
mechanism
catalysis
(beta/alpha)(8) barrel
glycoside hydrolase family 67
Pseudomonas
FLUORESCENS SUBSP CELLULOSA
SEQUENCE-BASED CLASSIFICATION
HYDROLASE FAMILY 10
CELLVIBRIO-MIXTUS
BINDING DOMAINS
ACTIVE-SITE
XYLANASE
SUBSTRATE
REFINEMENT
ENZYME

Cite this

@article{b516fbdc35fa4e418e8c018bd77fa7b3,

title = "The structural basis for catalysis and specificity of the Pseudomonas cellulosa alpha-glucuronidase, GlcA67A",

abstract = "alpha-glucuronidases, components of an ensemble of enzymes central to the recycling of photosynthetic biomass, remove the alpha-1,2 linked 4-O-methyl glucuronic acid from xylans. The structure of the alpha-glucuronidase, GlcA67A, from Pseudomonas cellulosa reveals three domains, the central of which is a (beta/alpha)(B) barrel housing the catalytic apparatus. Complexes of the enzyme with the individual reaction products, either xylobiose or glucuronic acid, and the ternary complex of both glucuronic acid and xylotriose reveal a {"}blind{"} pocket which selects for short decorated xylooligosaccharides substituted with the uronic acid at their nonreducing end, consistent with kinetic data. The catalytic center reveals a constellation of carboxylates; Glu292 is poised to provide protonic assistance to leaving group departure with Glu393 and Asp365 both appropriately positioned to provide base-catalyzed assistance for inverting nucleophilic attack by water.",

keywords = "glucuronidase, mechanism, catalysis, (beta/alpha)(8) barrel, glycoside hydrolase family 67, Pseudomonas, FLUORESCENS SUBSP CELLULOSA, SEQUENCE-BASED CLASSIFICATION, HYDROLASE FAMILY 10, CELLVIBRIO-MIXTUS, BINDING DOMAINS, ACTIVE-SITE, XYLANASE, SUBSTRATE, REFINEMENT, ENZYME",

author = "D Nurizzo and T Nagy and Gilbert, {H J} and Davies, {G J}",

year = "2002",

month = apr,

language = "English",

volume = "10",

pages = "547--556",

journal = "Structure",

issn = "0969-2126",

publisher = "Cell Press",

number = "4",

}

TY - JOUR

T1 - The structural basis for catalysis and specificity of the Pseudomonas cellulosa alpha-glucuronidase, GlcA67A

AU - Nurizzo, D

AU - Nagy, T

AU - Gilbert, H J

AU - Davies, G J

PY - 2002/4

Y1 - 2002/4

N2 - alpha-glucuronidases, components of an ensemble of enzymes central to the recycling of photosynthetic biomass, remove the alpha-1,2 linked 4-O-methyl glucuronic acid from xylans. The structure of the alpha-glucuronidase, GlcA67A, from Pseudomonas cellulosa reveals three domains, the central of which is a (beta/alpha)(B) barrel housing the catalytic apparatus. Complexes of the enzyme with the individual reaction products, either xylobiose or glucuronic acid, and the ternary complex of both glucuronic acid and xylotriose reveal a "blind" pocket which selects for short decorated xylooligosaccharides substituted with the uronic acid at their nonreducing end, consistent with kinetic data. The catalytic center reveals a constellation of carboxylates; Glu292 is poised to provide protonic assistance to leaving group departure with Glu393 and Asp365 both appropriately positioned to provide base-catalyzed assistance for inverting nucleophilic attack by water.

AB - alpha-glucuronidases, components of an ensemble of enzymes central to the recycling of photosynthetic biomass, remove the alpha-1,2 linked 4-O-methyl glucuronic acid from xylans. The structure of the alpha-glucuronidase, GlcA67A, from Pseudomonas cellulosa reveals three domains, the central of which is a (beta/alpha)(B) barrel housing the catalytic apparatus. Complexes of the enzyme with the individual reaction products, either xylobiose or glucuronic acid, and the ternary complex of both glucuronic acid and xylotriose reveal a "blind" pocket which selects for short decorated xylooligosaccharides substituted with the uronic acid at their nonreducing end, consistent with kinetic data. The catalytic center reveals a constellation of carboxylates; Glu292 is poised to provide protonic assistance to leaving group departure with Glu393 and Asp365 both appropriately positioned to provide base-catalyzed assistance for inverting nucleophilic attack by water.

KW - glucuronidase

KW - mechanism

KW - catalysis

KW - (beta/alpha)(8) barrel

KW - glycoside hydrolase family 67

KW - Pseudomonas

KW - FLUORESCENS SUBSP CELLULOSA

KW - SEQUENCE-BASED CLASSIFICATION

KW - HYDROLASE FAMILY 10

KW - CELLVIBRIO-MIXTUS

KW - BINDING DOMAINS

KW - ACTIVE-SITE

KW - XYLANASE

KW - SUBSTRATE

KW - REFINEMENT

KW - ENZYME

M3 - Article

SN - 0969-2126

VL - 10

SP - 547

EP - 556

JO - Structure

JF - Structure

IS - 4

ER -