In this study, a highly specific ratiometric two-photon fluorescent probe GP-BAN was developed and well characterized to monitor dipeptidyl peptidase IV in plasma and living systems. GP-BAN was designed on the basis of the catalytic properties and substrate preference of DPP-IV, and it could be readily hydrolyzed upon addition of DPP-IV under physiological conditions. Both reaction phenotyping and inhibition assays demonstrated that GP-BAN displayed good reactivity and high selectivity towards DPP-IV over other human serine hydrolases including FAP, DPP-VIII, and DPP-IX. The probe was successfully used to monitor the real activities of DPP-IV in complex biological systems including diluted plasma, while it could be used for high throughput screening of DPP-W inhibitors by using human plasma or tissue preparations as enzyme sources. As a two photon fluorescent probe, GP-BAN was also successfully used for two-photon imaging of endogenous DPP-IV in living cells and tissues, and showed high ratiometric imaging resolution and deep-tissue penetration ability. Taken together, a ratiometric two-photon fluorescent probe GP-BAN was developed and well-characterized for highly selective and sensitive detection of DPP-IV in complex biological systems, which could serve as a promising imaging tool to explore the biological functions and physiological roles of this key enzyme in living systems.
[1]Chinese Acad Sci, Dalian Inst Chem Phys, Dalian 116023, Peoples R China.
[2]Shanghai Univ Tradit Chinese Med, Shanghai, Peoples R China.
[3]Dalian Med Univ, Dalian 116044, Peoples R China.
[4]Wuhan Univ, Key Lab Analyt Chem Biol & Med, Minist Educ, Coll Chem & Mol Sci, Wuhan, Peoples R China.
(8.0+极速模式)