Literature summary for 2.4.99.18 extracted from

Maita, N.; Nyirenda, J.; Igura, M.; Kamishikiryo, J.; Kohda, D.
Comparative structural biology of eubacterial and archaeal oligosaccharyltransferases (2010), J. Biol. Chem., 285, 4941-4950.

Search for more literature for 2.4.99.18

Cloned(Commentary)

Cloned (Comment)	Organism
PCR-amplification fused to GST, expressed in Escherichia coli BL21 (DE3)pLysS	Campylobacter jejuni
the expression plasmid encoding the C-terminal soluble domain (residues 428713) is constructed by inserting a PCR product from genomic DNA (ATCC700819D) into the pGEX-6P-1 vector (GE Healthcare). The GST-fused sPglB protein is expressed by the addition of isopropyl-beta-D-thiogalactopyranoside at 37°C in the Escherichia coli BL21(DE3)pLysS strain. A structure-aided multiple sequence alignment of the STT3/PglB/AglB protein family reveals three types of oligosaccharyltransferases catalytic centers. This novel classification will provide a useful framework for understanding the enzymatic properties of the oligosaccharyltransferases enzymes from eukarya, archaea, and bacteria	Campylobacter jejuni

Cloned (Comment)

Organism

PCR-amplification fused to GST, expressed in Escherichia coli BL21 (DE3)pLysS

Campylobacter jejuni

the expression plasmid encoding the C-terminal soluble domain (residues 428713) is constructed by inserting a PCR product from genomic DNA (ATCC700819D) into the pGEX-6P-1 vector (GE Healthcare). The GST-fused sPglB protein is expressed by the addition of isopropyl-beta-D-thiogalactopyranoside at 37°C in the Escherichia coli BL21(DE3)pLysS strain. A structure-aided multiple sequence alignment of the STT3/PglB/AglB protein family reveals three types of oligosaccharyltransferases catalytic centers. This novel classification will provide a useful framework for understanding the enzymatic properties of the oligosaccharyltransferases enzymes from eukarya, archaea, and bacteria

Campylobacter jejuni

Crystallization (Commentary)

Crystallization (Comment)	Organism
2.8 A resolution crystal structure of the C-terminal globular domain, hanging drop method	Campylobacter jejuni
hanging drop crystallization with 10 mM Tris-HCl, pH 8.0, and reservoir solution (0.1 M sodium cacodylate, pH 6.5, 18% polyethylene glycol 8000, 0.2 M calcium acetate, 20°C, cryoprotection with 0.1 M MES, pH 6.5, polyethylene glycol 8000, and 0.2 M calcium acetate)	Campylobacter jejuni

Protein Variants

Protein Variants	Comment	Organism
D54A	XXD motif mutation, strongly reduced activity compared to wild-type	Campylobacter jejuni
I571A	MI motif mutation, strongly reduced activity compared to wild-type	Campylobacter jejuni
L570A	MI motif mutation, similar activity as wild-type	Campylobacter jejuni
M568A	MI motif mutation, similar activity as wild-type	Campylobacter jejuni
N53A	XXD motif mutation, about 40% activity remains compared to wild-type	Campylobacter jejuni
S52D	XXD motif mutation, strongly reduced activity compared to wild-type	Campylobacter jejuni
S52E	XXD motif mutation, strongly reduced activity compared to wild-type	Campylobacter jejuni
S569A	MI motif mutation, similar activity as wild-type	Campylobacter jejuni

Localization

Localization	Comment	Organism	GeneOntology No.	Textmining
membrane	-	Campylobacter jejuni	16020	-

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
dolichyl diphosphooligosaccharide + protein L-asparagine	Campylobacter jejuni	-	dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine	-	?
dolichyl diphosphooligosaccharide + protein L-asparagine	Campylobacter jejuni RM1221	-	dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine	-	?

Organism

Organism	UniProt	Comment	Textmining
Campylobacter jejuni	Q5HTX9	-	-

Purification (Commentary)

Purification (Comment)	Organism
sonication, centrifugation, supernatant absorbed to glutathione-Sepharose 4B resin, elution, concentration, reductive methylation, gel filtration with Superdex75 column, followed by anion exchange chromatography with a Resource Q column	Campylobacter jejuni

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
crude lipid-linked oligosaccharide donors from Campylobacter jejuni + carboxytetramethylrhodamine-Ala-Asp-Gln-Asn-Ala-Thr-Tyr-Lys	37°C, 50 mM Tris-HCl, pH 7.5	Campylobacter jejuni	dolichyl disphosphate + carboxytetramethylrhodamine-Ala-(oligosaccharidyl) Asp-Gln-Asp-Ala-Thr-Tyr	-	?
crude lipid-linked oligosaccharide donors from Campylobacter jejuni + carboxytetramethylrhodamine-Ala-Asp-Gln-Asn-Ala-Thr-Tyr-Lys	37°C, 50 mM Tris-HCl, pH 7.5	Campylobacter jejuni RM1221	dolichyl disphosphate + carboxytetramethylrhodamine-Ala-(oligosaccharidyl) Asp-Gln-Asp-Ala-Thr-Tyr	-	?
dolichyl diphosphooligosaccharide + protein L-asparagine	-	Campylobacter jejuni	dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine	-	?
dolichyl diphosphooligosaccharide + protein L-asparagine	-	Campylobacter jejuni RM1221	dolichyl disphosphate + protein with oligosaccharide attached to protein L-asparagine	-	?

Synonyms

Synonyms	Comment	Organism
oligosaccharyltransferase	-	Campylobacter jejuni
OST	-	Campylobacter jejuni
PglB	-	Campylobacter jejuni
PglB	single-subunit membrane protein	Campylobacter jejuni

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
7.5	-	assay at	Campylobacter jejuni