PUBLICATION

Structure of the polyisoprenyl-phosphate glycosyltransferase GtrB and insights into the mechanism of catalysis

Authors

Ardiccioni, C., Clarke, O.B., Tomasek, D., Issa, H.A., von Alpen, D.C., Pond, H.L., Banerjee, S., Rajashankar, K.R., Liu, Q., Guan, Z., Li, C., Kloss, B., Bruni, R., Kloppmann, E., Rost, B., Manzini, M.C., Shapiro, L., Mancia, F.

ID

ZDB-PUB-160106-5

Date

2016

Source

Nature communications 7: 10175 (Journal)

Registered Authors

Keywords

Biocatalysis, Structural biology, Transferases

MeSH Terms

Models, Molecular
Mannosyltransferases/genetics
Mannosyltransferases/metabolism
Animals, Genetically Modified
Zebrafish
Animals
Glycosyltransferases/genetics
Glycosyltransferases/metabolism*
Humans
Synechocystis/enzymology*
Gene Expression Regulation, Bacterial/physiology*
Protein Conformation
Gene Expression Regulation, Enzymologic/physiology*

(all 13)

PubMed

26729507 Full text @ Nat. Commun.

Abstract

The attachment of a sugar to a hydrophobic polyisoprenyl carrier is the first step for all extracellular glycosylation processes. The enzymes that perform these reactions, polyisoprenyl-glycosyltransferases (PI-GTs) include dolichol phosphate mannose synthase (DPMS), which generates the mannose donor for glycosylation in the endoplasmic reticulum. Here we report the 3.0Å resolution crystal structure of GtrB, a glucose-specific PI-GT from Synechocystis, showing a tetramer in which each protomer contributes two helices to a membrane-spanning bundle. The active site is 15 Å from the membrane, raising the question of how water-soluble and membrane-embedded substrates are brought into apposition for catalysis. A conserved juxtamembrane domain harbours disease mutations, which compromised activity in GtrB in vitro and in human DPM1 tested in zebrafish. We hypothesize a role of this domain in shielding the polyisoprenyl-phosphate for transport to the active site. Our results reveal the basis of PI-GT function, and provide a potential molecular explanation for DPM1-related disease.

Genes / Markers

Figures

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Expression

Phenotype

Fish	Conditions	Stage	Phenotype	Figure
AB + MO3-dpm1	standard conditions	Long-pec	blood accumulation caudal fin distal region, abnormal	Fig. S15
AB + MO3-dpm1	standard conditions	Long-pec	caudal artery fused with caudal vein, abnormal	Fig. S15
AB + MO3-dpm1	standard conditions	Long-pec	caudal artery increased size, abnormal	Fig. S15
AB + MO3-dpm1	standard conditions	Long-pec	caudal fin kinked, abnormal	Fig. 4
AB + MO3-dpm1	standard conditions	Long-pec	caudal vein increased size, abnormal	Fig. S15
AB + MO3-dpm1	standard conditions	Long-pec	eye decreased size, abnormal	Fig. 4
AB + MO3-dpm1	standard conditions	Long-pec	eye decreased size, abnormal	Fig. S15
AB + MO3-dpm1	standard conditions	Long-pec	head decreased size, abnormal	Fig. 4
AB + MO3-dpm1	standard conditions	Long-pec	head decreased size, abnormal	Fig. S15
AB + MO3-dpm1	standard conditions	Long-pec	vasculature morphology, abnormal	Fig. 4

1 - 10 of 20

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Mutations / Transgenics

Human Disease / Model

Human Disease	Fish	Conditions	Evidence
congenital disorder of glycosylation type I	AB + MO4-dpm1	standard conditions	IC
congenital disorder of glycosylation type I	AB + MO3-dpm1	standard conditions	IC

1 - 2 of 2

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Sequence Targeting Reagents

Target	Reagent	Reagent Type
dpm1	MO3-dpm1	MRPHLNO
dpm1	MO4-dpm1	MRPHLNO

Fish

Fish
AB + MO3-dpm1
AB + MO4-dpm1

Antibodies

Orthology

Engineered Foreign Genes

Mapping

Ardiccioni et al., 2016 ZDB-PUB-160106-5 PMID:26729507

Structure of the polyisoprenyl-phosphate glycosyltransferase GtrB and insights into the mechanism of catalysis

Ardiccioni et al., 2016

ZDB-PUB-160106-5
PMID:26729507