p-Guanidinobenzoic acid esters of fluorescein as active-site titrants of serine proteases
- 1 December 1982
- journal article
- research article
- Published by American Chemical Society (ACS) in Journal of the American Chemical Society
- Vol. 104 (25), 7299-7306
- https://doi.org/10.1021/ja00389a065
Abstract
Two active-site titrants of serine proteases, fluorescein mono-p-guanidinobenzoate hydrochloride (FMGB.cntdot.HCl) and fluorescein di-p-guanidinobenzoate dihydrochloride (FDGB.cntdot.2HCl), were synthesized, purified, and chemically and enzymatically characterized. Electronic absorption and fluorescence emission spectra, fluorescence lifetimes and quantum yields, solubilities, and rates of spontaneous hydrolysis at pH 7-10 are reported. Macroscopic and microscopic kinetic constants for interaction of FMGB.cntdot.HCl with trypsin, urokinase, plasmin, and thrombin were determined. FMGB.cntdot.HCl, which rapidly releases fluorescein upon formation of a stable acyl-enzyme intermediate with trypsin and other trypsin-like enzymes, is the most sensitive active-site titrant for serine proteases yet described.This publication has 9 references indexed in Scilit:
- A quantitative assay for the activation of plasminogen by transformed cells in situ and by urokinaseBiochemistry, 1981
- Synthesis and characterization of a new fluorogenic active-site titrant of serine proteasesBiochemistry, 1981
- Sensitive assay for plasminogen activator of transformed cells.Proceedings of the National Academy of Sciences of the United States of America, 1980
- Cleavage of the Escherichia coli lexA protein by the recA protease.Proceedings of the National Academy of Sciences of the United States of America, 1980
- Plasminogen activator of islets of Langerhans: modulation by glucose and correlation with insulin production.Proceedings of the National Academy of Sciences, 1980
- Trypsin liberates an arginine vasopressin-like peptide and neurophysin from a Mr 20,000 putative common precursor.Proceedings of the National Academy of Sciences of the United States of America, 1979
- Plasminogen activator in early embryogenesis: Enzyme production by trophoblast and parietal endodermCell, 1976
- Studies on the role of plasminogen activator in ovulation. In vitro response of granulosa cells to gonadotropins, cyclic nucleotides, and prostaglandins.Journal of Biological Chemistry, 1976
- Lipolysis of fluorescein and eosin esters. Kinetics of hydrolysisAnalytical Biochemistry, 1966