Substrate optimization for monitoring cathepsin C activity in live cells

详细信息    查看全文

• 作者：Jun Li ; H. Michael Petrassi ; Christine Tumanut ; Brian T. Masick ; Christopher Trussell ; Jennifer L. Harris
• 关键词：Cathepsin ; Imaging ; Rhodamine ; Activity-based probe
• 刊名：Bioorganic and Medicinal Chemistry
• 出版年：2009
• 出版时间：1 February 2009
• 年：2009
• 卷：17
• 期：3
• 页码：1064-1070
• 全文大小：283 K

文摘

A series of peptidic fluorogenic substrates were synthesized to develop a flow cytometry assay (FACS) to monitor the proteolytic activity of cathepsin C in live cells. Of the 16 substrates tested, (NH₂-aminobutyric-homophenylalanine)₂-rhodamine demonstrated the best reactivity and selectivity profile in the FACS assay using the B721 human B-lymphoblastoid cell line. The resulting FACS assay was validated through correlation of the IC₅₀ values with a competitive radiolabeling assay against a series of small molecule inhibitors of cathepsin C.