Literature summary for 2.7.8.43 extracted from

Fage, C.D.; Brown, D.B.; Boll, J.M.; Keatinge-Clay, A.T.; Trent, M.S.
Crystallographic study of the phosphoethanolamine transferase EptC required for polymyxin resistance and motility in Campylobacter jejuni (2014), Acta Crystallogr. Sect. D, 70, 2730-2739.

Cloned(Commentary)

Cloned (Comment)	Organism
gene eptC, recombinant expression of C-terminally Gly6-His8-tagged enzyme in Escherichia coli strain BL21(DE3), recombinant expression of His-tagged enzyme as selenomethionine-labeled enzyme	Campylobacter jejuni

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant His-tagged wild-type and selenomethionine-labeled enzyme EptC, sitting drop vapour diffusion method, mixing of 0.004 ml of 30 mg/ml protein in 25 mM NaCl, 10 mM HEPES, pH 7.5, with 0.001 ml of precipitant solution containing 200 mM diammonium phosphate, 15% w/v PEG 3350, 22°C, X-ray diffraction structure determination and analysis at 2.40-2.80 A resolution, molecular replacement	Campylobacter jejuni

Crystallization (Comment)

Organism

purified recombinant His-tagged wild-type and selenomethionine-labeled enzyme EptC, sitting drop vapour diffusion method, mixing of 0.004 ml of 30 mg/ml protein in 25 mM NaCl, 10 mM HEPES, pH 7.5, with 0.001 ml of precipitant solution containing 200 mM diammonium phosphate, 15% w/v PEG 3350, 22°C, X-ray diffraction structure determination and analysis at 2.40-2.80 A resolution, molecular replacement

Campylobacter jejuni

Protein Variants

Protein Variants	Comment	Organism
H358A	site-directed mutagenesis, mutation of a potential zinc binding residue	Campylobacter jejuni
H440A	site-directed mutagenesis, mutation of a potential zinc binding residue	Campylobacter jejuni
N308A	site-directed mutagenesis, mutation of a potential zinc binding residue	Campylobacter jejuni
S309A	site-directed mutagenesis, mutation of a potential zinc binding residue	Campylobacter jejuni
T266A	site-directed mutagenesis, mutation of a potential zinc binding residue	Campylobacter jejuni
T266S	site-directed mutagenesis, mutation of a potential zinc binding residue	Campylobacter jejuni

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
inner membrane	phosphoethanolamine modification occurs on the periplasmic side of the inner membrane, where enzyme EptC transfers head groups from the phospholipid phosphoethanolamine to target molecules. EptC is predicted to be anchored to the inner membrane by a five-helix transmembrane domain, with its sulfatase-like catalytic domain positioned in the periplasm	Campylobacter jejuni	-	-

Metals/Ions

Metals/Ions	Comment	Organism	Structure
Zn2+	zinc-binding residues of the phosphoenzyme intermediate, overview. The cEptC active site contains a tetrahedrally coordinated zinc ion and a putative nucleophilic threonine, Thr266, covalently bound to a phosphoryl group to form phosphothreonine	Campylobacter jejuni

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
36700	-	-	Campylobacter jejuni

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.
diacylphosphatidylethanolamine + flagellar rod protein FlgG	Campylobacter jejuni	-	?	-	?
diacylphosphatidylethanolamine + flagellar rod protein FlgG	Campylobacter jejuni 81-176	-	?	-	?
diacylphosphatidylethanolamine + lipid A	Campylobacter jejuni	-	diacylglycerol + lipid A 1-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	Campylobacter jejuni	-	diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	Campylobacter jejuni 81-176	-	diacylglycerol + lipid A 1-(2-aminoethyl diphosphate)	-	?
additional information	Campylobacter jejuni	pEtN transferase, EptC, modifies an array of cell-surface molecules and the N-linked glycans of numerous glycoproteins	?	-	?
additional information	Campylobacter jejuni 81-176	pEtN transferase, EptC, modifies an array of cell-surface molecules and the N-linked glycans of numerous glycoproteins	?	-	?

Organism

Organism	UniProt	Comment	Textmining
Campylobacter jejuni	A0A3Z8TVG8	-	-
Campylobacter jejuni 81-176	A0A3Z8TVG8	-	-

Posttranslational Modification

Posttranslational Modification	Comment	Organism
phosphoprotein	the cEptC active site contains a putative nucleophilic threonine, Thr266, covalently bound to a phosphoryl group to form phosphothreonine. Phospho-Thr266 mimics an enzyme reaction intermediate representing the first step in a two-step transferase reaction	Campylobacter jejuni

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and selenomethionine-labeled enzyme from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration	Campylobacter jejuni

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
diacylphosphatidylethanolamine + flagellar rod protein FlgG	-	Campylobacter jejuni	?	-	?
diacylphosphatidylethanolamine + flagellar rod protein FlgG	EptC transfers phosphoethanolamine from the head groups of phosphatidylethanolamine onto Thr75 of FlgG proteins	Campylobacter jejuni	?	-	?
diacylphosphatidylethanolamine + flagellar rod protein FlgG	-	Campylobacter jejuni 81-176	?	-	?
diacylphosphatidylethanolamine + flagellar rod protein FlgG	EptC transfers phosphoethanolamine from the head groups of phosphatidylethanolamine onto Thr75 of FlgG proteins	Campylobacter jejuni 81-176	?	-	?
diacylphosphatidylethanolamine + lipid A	-	Campylobacter jejuni	diacylglycerol + lipid A 1-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	EptC transfers phosphoethanolamine from the head groups of phosphatidylethanolamine onto the LOS core and phosphates of lipid A	Campylobacter jejuni	diacylglycerol + lipid A 1-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	-	Campylobacter jejuni 81-176	diacylglycerol + lipid A 1-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	EptC transfers phosphoethanolamine from the head groups of phosphatidylethanolamine onto the LOS core and phosphates of lipid A	Campylobacter jejuni 81-176	diacylglycerol + lipid A 1-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	-	Campylobacter jejuni	diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate)	-	?
diacylphosphatidylethanolamine + lipid A	EptC transfers phosphoethanolamine from the head groups of phosphatidylethanolamine onto the LOS core and phosphates of lipid A	Campylobacter jejuni	diacylglycerol + lipid A 4'-(2-aminoethyl diphosphate)	-	?
additional information	pEtN transferase, EptC, modifies an array of cell-surface molecules and the N-linked glycans of numerous glycoproteins	Campylobacter jejuni	?	-	?
additional information	pEtN transferase, EptC, modifies an array of cell-surface molecules and the N-linked glycans of numerous glycoproteins	Campylobacter jejuni 81-176	?	-	?

Subunits

Subunits	Comment	Organism
?	x * 36700, recombinant His-tagged wild-type, SDS-PAGE	Campylobacter jejuni
More	enzyme structure analysis, overview	Campylobacter jejuni

Synonyms

Synonyms	Comment	Organism
Cj0256	locus name	Campylobacter jejuni
EptC	-	Campylobacter jejuni
pEtN transferase	-	Campylobacter jejuni
phosphoethanolamine transferase	-	Campylobacter jejuni

General Information

General Information	Comment	Organism
evolution	the eptC gene (locus tag Cj0256) is clustered in a family of inner-membrane metalloenzymes (COG2194) containing a fivehelix transmembrane domain and a periplasmic catalytic domain that is currently grouped in the sulfatase family	Campylobacter jejuni
additional information	identification of identify zinc-ligand residues, a putative nucleophile and conserved active-site residues required for in vivo activity from the crystal structure, active-site architecture of cEptC, overview	Campylobacter jejuni
physiological function	the foodborne enteric pathogen Campylobacter jejuni decorates a variety of its cell-surface structures with phosphoethanolamine. Modifying lipid A with phosphoethanolamine promotes cationic antimicrobial peptide resistance. Modifications of the Campylobacter jejuni surface structures with phosphoethanolamine promote flagellar assembly, motility, cationic antimicrobial peptide resistance and host intestinal colonization	Campylobacter jejuni