Literature summary for 2.7.7.62 extracted from

Otte, M.M.; Woodson, J.D.; Escalante-Semerena, J.C.
The thiamine kinase (YcfN) enzyme plays a minor but significant role in cobinamide salvaging in Salmonella enterica (2007), J. Bacteriol., 189, 7310-7315.

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Cloned(Commentary)

Cloned (Comment)	Organism
in pET-16b (pCLK740) for expression with N-terminal deca-His-tag in Salmonella enterica serovar Typhimurium LT2, strain JE8407 lacking endogenous CobU and YcfN	Escherichia coli

General Stability

General Stability	Organism
no soluble N-terminal His-tagged protein detectable after immobilized metal affinity chromatography from over-expressing Salmonella enterica serovar Typhimurium LT2	Escherichia coli

Molecular Weight [Da]

Molecular Weight [Da]	Molecular Weight Maximum [Da]	Comment	Organism
34000	-	SDS-PAGE	Escherichia coli

Organism

Organism	UniProt	Comment	Textmining
Escherichia coli	-	-	-
Salmonella enterica	-	serovar typhimurium LT2	-

Purification (Commentary)

Purification (Comment)	Organism
tandem hydrophobic interaction chromatography on two HiTrap Phenyl FastFlow columns, dialysation, and dye ligand chromatography on HiTrap Blue HP column (elution with 2 M NaCl)	Salmonella enterica

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
adenosylcobinamide-GDP + diphosphate	2 mM ATP, 5 mM MgCl2, 16 h, pH 8, 37°C	Salmonella enterica	GTP + adenosylcobinamide phosphate	reaction stop by 25 mM KCN and 10 min heating at 80°C, analysis by mass spectrometry	?
adenosylcobinamide-GDP + diphosphate	over-expressing cell extracts, 2 mM ATP, 5 mM MgCl2, 16 h, pH 8, 37°C	Escherichia coli	GTP + adenosylcobinamide phosphate	reaction stop by 25 mM KCN and 10 min heating at 80°C, analysis by mass spectrometry	?

Synonyms

Synonyms	Comment	Organism
CobU	-	Salmonella enterica
GTP: adenosylcobinamide-phosphate guanylyltransferase	GTP: AdoCbi-P guanylyltransferase	Salmonella enterica
NTP: adenosylcobinamide kinase	NTP: AdoCbi kinase	Salmonella enterica
thiamine kinase	-	Escherichia coli
YcfN	-	Escherichia coli

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Release 2023.1 (January 2023)