The thiamine kinase (YcfN) enzyme plays a minor but significant role in cobinamide salvaging in Salmonella enterica.
Cobinamide (Cbi) salvaging is impaired, but not abolished, in a Salmonella enterica strain lacking a functional cobU gene. CobU is a bifunctional enzyme (NTP:adenosylcobinamide [NTP:AdoCbi] kinase, GTP:adenosylcobinamide-phosphate [GTP:AdoCbi-P] guanylyltransferase) whose AdoCbi kinase activity is necessary for Cbi salvaging in this bacterium. Inactivation of the ycfN gene in a DeltacobU strain abrogated Cbi salvaging. Introduction of a plasmid carrying the ycfN(+) allele into a DeltacobU DeltaycfN strain substantially restored Cbi salvaging. Mass spectrometry data indicate that when YcfN-enriched cell extracts were incubated with AdoCbi and ATP, the product of the reaction was AdoCbi-P. Results from bioassays confirmed that YcfN converted AdoCbi to AdoCbi-P in an ATP-dependent manner. YcfN is a good example of enzymes that are used by the cell in multiple pathways to ensure the salvaging of valuable precursors.
Otte
Michele M
MM
Department of Bacteriology, University of Wisconsin-Madison, 144A Enzyme Institute, Madison, WI 53726-4087, USA.
Woodson
Jesse D
JD
Escalante-Semerena
Jorge C
JC
D000255:Adenosine Triphosphate / Q000378:metabolism; D003038:Cobamides / Q000378:metabolism*; D003600:Cytosol / Q000737:chemistry; D017353:Gene Deletion; D005816:Genetic Complementation Test; D013058:Mass Spectrometry; D016254:Mutagenesis, Insertional; D010755:Organophosphates / Q000378:metabolism; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D019779:Salmonella enterica / Q000201:enzymology* / Q000235:genetics / Q000502:physiology
D003038:Cobamides; D010755:Organophosphates; C524259:adenosylcobinamide phosphate; C414173:adensoylcobinamide; C017142:cobinamide; D000255:Adenosine Triphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); C035182:thiamin kinase
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