Abstract
Objectives: The objective of this work was to demonstrate the utility of luminescence from lanthanides bound to a mutant of the Ca 2+ binding protein, oncomodulin, to monitor protease activity. Design and Methods: A mutant of oncomodulin with a cysteine residue at position 57 located in the CD binding loop was conjugated to a salicylic acid group. The luminescence of Tb 3+ resulting from electronic energy transfer from the salicylic acid group was monitored using time resolved lanthanide luminescence in the presence of proteolytic enzymes. Results: Low detection limits for subtilisin (150 pg), chrymotrypsin (2.5 ng), cathepsin B (3.5 ng), and HIV-1 protease (25 ng) were found. Conclusion: The simplicity of the assay coupled with its high level of sensitivity make it useful for the detection of proteases at very low concentrations.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.