A new type of ultrasensitive bioluminogenic enzyme substrates. I. Enzyme substrates with D-luciferin as leaving group.
Derivatives of D-luciferin, D-luciferin methyl ester, D-luciferin O-sulfate, D-luciferin O-phosphate, D-luciferyl-L-N alpha-arginine and D-luciferyl-L-phenylalanine were used as highly sensitive substrates for carboxylic esterase, arylsulfatase, alkaline phosphatase and carboxypeptidases A, B and N. Enzymatic cleavage of the compounds by enzymes leading to the release of D-luciferin was demonstrated. Kinetic constants have been determined for D-luciferin methyl ester and carboxylic esterase, for D-luciferin O-sulfate and arylsulfatase, for D-luciferin O-phosphate and alkaline phosphatase, for D-luciferyl-L-phenylalanine and carboxypeptidase A, and for carboxypeptidases B and N and D-luciferyl-L-N alpha-arginine. All compounds proved to be highly sensitive substrates for the respective enzymes, permitting a limit of detection for enzymes between 10 and 500 fg per assay.
Miska
W
W
Dermatologische Universitätsklinik, München.
Geiger
R
R
D000469:Alkaline Phosphatase / Q000378:metabolism*; D001192:Arylsulfatases / Q000378:metabolism*; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism*; D043424:Carboxypeptidase B; D002268:Carboxypeptidases / Q000378:metabolism*; D043422:Carboxypeptidases A; D049454:Firefly Luciferin / Q000031:analogs & derivatives*; D007700:Kinetics; D008156:Luciferases / Q000378:metabolism*; D008163:Luminescent Measurements; D001121:Lysine Carboxypeptidase / Q000378:metabolism; D008722:Methods; D013379:Substrate Specificity; D013429:Sulfatases / Q000378:metabolism*
D049454:Firefly Luciferin; D008156:Luciferases; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D000469:Alkaline Phosphatase; D013429:Sulfatases; D001192:Arylsulfatases; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D043424:Carboxypeptidase B; D001121:Lysine Carboxypeptidase