Enzymatic cleavage and subsequent facile intramolecular transcyclization for in situ fluorescence detection of γ-glutamyltranspetidase activities

Hongjuan Tong, Yongjun Zheng, Li Zhou, Xiangmin Li, Rui Qian, Rui Wang, Jianhong Zhao, Kaiyan Lou, Wei Wang

Research output: Contribution to journalArticlepeer-review

55 Scopus citations

Abstract

γ-Glutamyltranspetidase (GGT) is a cell-membrane-bound enzyme which selectively catalyzes cleavage of the γ-glutamyl bond of glutathione (GSH). It has been identified to be overexpressed in a number of malignant tumor cells. Therefore, fluorescent probes for fast and selective detection of GGT activities are greatly needed. However, the majority of currently available GGT fluorescent probes based on direct conjugation of a γ-glutamyl group to a specific fluorophore generally has slow enzymatic kinetics due to bulky fluorophore too close to the enzyme's active site. Moreover, the uncaged fluorophore with a free amine group might undergo oxidation or other enzymatic transformation and resulted in a complicated timedependent fluorescence response. Herein, we reported design of a novel fluorescent GGT probe NM-GSH (2), which incorporated a fast intramolecular transcyclization cascade for rapid detection of GGT activities after enzymatic cleavage of the γ-glutamyl group. This design strategy allows introduction of bulky 1,8-naphthalimide fluorophore with improved enzymatic kinetics and lowered detection limit. The transcyclized product 4 gives more than 200-fold fluorescence increment. The probe NM-GSH showed both good selectivity and fast detection of GGT activities with the detection limit as low as 0.21 mU/mL. In addition, the fluorescent product 4 contains no free amine group and is more stable for detection. Most importantly, cell imaging studies showed that the transcyclized product 4 was enriched in lysosomes for selectively lighting up GGT-overexpressed ovarian cancer cells (OVCAR5) but not normal cells (HUVEC), indicating NMGSH's potentials as an imaging agent in cancer diagnosis and treatment.

Original languageEnglish (US)
Pages (from-to)10816-10820
Number of pages5
JournalAnalytical Chemistry
Volume88
Issue number22
DOIs
StatePublished - Nov 15 2016
Externally publishedYes

ASJC Scopus subject areas

  • Analytical Chemistry

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