Active-site labelling of triose phosphate isomerase. The reaction of bromohydroxyacetone phosphate with a unique glutamic acid residue and the migration of the label to tyrosine.
Triose phosphate isomerase from chicken muscle reacts stoicheiometrically with the active-site-directed irreversible inhibitor bromohydroxyacetone phosphate with concomitant loss of all catalytic activity. The primary site of attachment has been shown to be a unique glutamic acid residue in the sequence Ala-Tyr-Glu-Pro-Val-Trp. Unless the inhibitor-enzyme bond is stabilized by reduction of the C-2 carbonyl group with borohydride, the phosphate group is lost and the label migrates to the adjacent tyrosine residue. It is suggested that the gamma-carboxylate group of the glutamic acid residue may be the base responsible for primary proton abstraction from substrate in the catalysis. The failure of this reagent specifically to inactivate either muscle or yeast aldolase, and the use of the reagent in preparing isomerase-free glycolytic enzymes, is discussed.
D000096:Acetone / Q000031:analogs & derivatives; D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D001966:Bromine; D002238:Carbohydrate Epimerases; D002247:Carbon Isotopes; D002645:Chickens; D005634:Fructose-Bisphosphate Aldolase; D005971:Glutamates; D006016:Glycolates; D009132:Muscles / Q000201:enzymology; D009943:Organophosphorus Compounds; D010455:Peptides / Q000302:isolation & purification; D010759:Phosphorus Isotopes; D013056:Spectrophotometry, Ultraviolet; D014306:Trioses; D014443:Tyrosine
D002247:Carbon Isotopes; D005971:Glutamates; D006016:Glycolates; D009943:Organophosphorus Compounds; D010455:Peptides; D010759:Phosphorus Isotopes; D014306:Trioses; D000096:Acetone; C002417:bromohydroxyacetone phosphate; D014443:Tyrosine; D005634:Fructose-Bisphosphate Aldolase; D002238:Carbohydrate Epimerases; D001966:Bromine