Structure and mechanism of a bacterial beta-glucosaminidase having O-GlcNAcase activity

R J  Dennis; E J  Taylor; M S  Macauley; K A  Stubbs; J P  Turkenburg; S J  Hart; G N  Black; D J  Vocadlo; G J  Davies

doi:10.1038/nsmb1079

Structure and mechanism of a bacterial beta-glucosaminidase having O-GlcNAcase activity

R J Dennis, E J Taylor, M S Macauley, K A Stubbs, J P Turkenburg, S J Hart, G N Black, D J Vocadlo, G J Davies

Chemistry

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

Abstract

O-GlcNAc is an abundant post-translational modification of serine and threonine residues of nucleocytoplasmic proteins. This modification, found only within higher eukaryotes, is a dynamic modification that is often reciprocal to phosphorylation. In a manner analogous to phosphatases, a glycoside hydrolase termed O-GlcNAcase cleaves O-GlcNAc from modified proteins. Enzymes with high sequence similarity to human O-GlcNAcase are also found in human pathogens and symbionts. We report the three-dimensional structure of O-GlcNAcase from the human gut symbiont Bacteroides thetaiotaomicron both in its native form and in complex with a mimic of the reaction intermediate. Mutagenesis and kinetics studies show that the bacterial enzyme, very similarly to its human counterpart, operates via an unusual 'substrate-assisted' catalytic mechanism, which will inform the rational design of enzyme inhibitors.

Original language	English
Pages (from-to)	365-371
Number of pages	7
Journal	Nature Structural & Molecular Biology
Volume	13
Issue number	13
DOIs	https://doi.org/10.1038/nsmb1079
Publication status	Published - 26 Mar 2006

Keywords

SUBSTRATE-ASSISTED CATALYSIS
TAY-SACHS-DISEASE
N-ACETYLGLUCOSAMINE
CYTOSOLIC PROTEINS
LINKED GLCNAC
NUCLEOCYTOPLASMIC PROTEINS
TETRATRICOPEPTIDE REPEATS
CRYSTAL-STRUCTURE
GLYCOSYLATION
TRANSFERASE

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10.1038/nsmb1079

http://dx.doi.org/10.1038/nsmb1079

Cite this

@article{a3dad0626ed44c95bbe729e0f6256d03,

title = "Structure and mechanism of a bacterial beta-glucosaminidase having O-GlcNAcase activity",

abstract = "O-GlcNAc is an abundant post-translational modification of serine and threonine residues of nucleocytoplasmic proteins. This modification, found only within higher eukaryotes, is a dynamic modification that is often reciprocal to phosphorylation. In a manner analogous to phosphatases, a glycoside hydrolase termed O-GlcNAcase cleaves O-GlcNAc from modified proteins. Enzymes with high sequence similarity to human O-GlcNAcase are also found in human pathogens and symbionts. We report the three-dimensional structure of O-GlcNAcase from the human gut symbiont Bacteroides thetaiotaomicron both in its native form and in complex with a mimic of the reaction intermediate. Mutagenesis and kinetics studies show that the bacterial enzyme, very similarly to its human counterpart, operates via an unusual 'substrate-assisted' catalytic mechanism, which will inform the rational design of enzyme inhibitors.",

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author = "Dennis, {R J} and Taylor, {E J} and Macauley, {M S} and Stubbs, {K A} and Turkenburg, {J P} and Hart, {S J} and Black, {G N} and Vocadlo, {D J} and Davies, {G J}",

year = "2006",

month = mar,

day = "26",

doi = "10.1038/nsmb1079",

language = "English",

volume = "13",

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journal = "Nature Structural & Molecular Biology",

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T1 - Structure and mechanism of a bacterial beta-glucosaminidase having O-GlcNAcase activity

AU - Dennis, R J

AU - Taylor, E J

AU - Macauley, M S

AU - Stubbs, K A

AU - Turkenburg, J P

AU - Hart, S J

AU - Black, G N

AU - Vocadlo, D J

AU - Davies, G J

PY - 2006/3/26

Y1 - 2006/3/26

N2 - O-GlcNAc is an abundant post-translational modification of serine and threonine residues of nucleocytoplasmic proteins. This modification, found only within higher eukaryotes, is a dynamic modification that is often reciprocal to phosphorylation. In a manner analogous to phosphatases, a glycoside hydrolase termed O-GlcNAcase cleaves O-GlcNAc from modified proteins. Enzymes with high sequence similarity to human O-GlcNAcase are also found in human pathogens and symbionts. We report the three-dimensional structure of O-GlcNAcase from the human gut symbiont Bacteroides thetaiotaomicron both in its native form and in complex with a mimic of the reaction intermediate. Mutagenesis and kinetics studies show that the bacterial enzyme, very similarly to its human counterpart, operates via an unusual 'substrate-assisted' catalytic mechanism, which will inform the rational design of enzyme inhibitors.

AB - O-GlcNAc is an abundant post-translational modification of serine and threonine residues of nucleocytoplasmic proteins. This modification, found only within higher eukaryotes, is a dynamic modification that is often reciprocal to phosphorylation. In a manner analogous to phosphatases, a glycoside hydrolase termed O-GlcNAcase cleaves O-GlcNAc from modified proteins. Enzymes with high sequence similarity to human O-GlcNAcase are also found in human pathogens and symbionts. We report the three-dimensional structure of O-GlcNAcase from the human gut symbiont Bacteroides thetaiotaomicron both in its native form and in complex with a mimic of the reaction intermediate. Mutagenesis and kinetics studies show that the bacterial enzyme, very similarly to its human counterpart, operates via an unusual 'substrate-assisted' catalytic mechanism, which will inform the rational design of enzyme inhibitors.

KW - SUBSTRATE-ASSISTED CATALYSIS

KW - TAY-SACHS-DISEASE

KW - N-ACETYLGLUCOSAMINE

KW - CYTOSOLIC PROTEINS

KW - LINKED GLCNAC

KW - NUCLEOCYTOPLASMIC PROTEINS

KW - TETRATRICOPEPTIDE REPEATS

KW - CRYSTAL-STRUCTURE

KW - GLYCOSYLATION

KW - TRANSFERASE

U2 - 10.1038/nsmb1079

DO - 10.1038/nsmb1079

M3 - Article

SN - 1545-9993

VL - 13

SP - 365

EP - 371

JO - Nature Structural & Molecular Biology

JF - Nature Structural & Molecular Biology

IS - 13

ER -