Bovine inositol monophosphatase: development of a continuous fluorescence assay of enzyme activity.
This paper describes a continuous assay for the enzyme inositol monophosphatase which has been developed using a new substrate, the fluorescent compound 4-methylumbelliferyl phosphate. The hydrolysis of the phosphate group from this compound can be readily detected by a resultant large red shift in the emission spectrum from 390-450 nm. The kinetic constants for the enzyme using this new substrate are described.
Gore
M G
MG
Dept. of Biochemistry, School of Biological Sciences, University of Southampton, UK.
Greasley
P J
PJ
Ragan
C I
CI
D000818:Animals; D002417:Cattle; D003511:Cyclohexanols / Q000494:pharmacology; D005453:Fluorescence; D006923:Hymecromone / Q000031:analogs & derivatives / Q000378:metabolism; D007700:Kinetics; D008094:Lithium / Q000494:pharmacology; D008274:Magnesium / Q000494:pharmacology; D009943:Organophosphorus Compounds / Q000494:pharmacology; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D013053:Spectrophotometry / Q000379:methods
D003511:Cyclohexanols; D009943:Organophosphorus Compounds; C069565:1-phosphoryloxy-2,4-dihydroxycyclohexane; C005359:4-methylumbelliferyl phosphate; D006923:Hymecromone; D008094:Lithium; D010744:Phosphoric Monoester Hydrolases; C021822:myo-inositol-1 (or 4)-monophosphatase; D008274:Magnesium