Rational Design of a Profluorescent Substrate for S-adenosylhomocysteine Hydrolase and its Applications in Bioimaging and Inhibitor Screening.
S-Adenosylhomocysteine hydrolase (SAHase) is a cellular enzyme that plays a key role in the methylation process, and a potential drug target in the discovery of antiviral and anticancer agents. There is increasing interest in determining its activity in the biological and clinical fields with chemosensors but with limited success so far. Herein, we designed and developed for the first time an off/on-type of fluorogenic substrate (NADE) that is directly responsive to SAHase activity. NADE used 1,8-naphthalimide as the signal reporter and adenosine (Ade) as the reaction center; removal of the Ade moiety enhanced the fluorescence by >10-fold. Kinetic study showed that NADE followed a non-Michaelis-Menten pattern that corresponded to the allosteric behavior of SAHase. NADE showed excellent selectivity and functioned efficiently in cells, allowing the microscopic imaging of SAHase activity. NADE can also be used to identify and measure the effectiveness of inhibitors in a markedly superior way. In a word, NADE would be broadly useful in clinical applications and academic studies.
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.|Graduate School of the Chinese Academy of Sciences , Beijing, P. R. China.
Yan
Y
Jia
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.
Peng
P
Li
The First Affiliated Hospital of Dalian Medical University , Dalian 116023, P. R. China.
Wei
W
Song
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.
Guangjiu
G
Zhao
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.
Daoyuan
D
Zheng
College of Pharmacy, Nankai University , Haihe Education Park, 38 Tongyan Road, Tianjin 300353, People's Republic of China.
Dongmei
D
Li
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.|Graduate School of the Chinese Academy of Sciences , Beijing, P. R. China.
Yanni
Y
Wang
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.
Jiayue
J
Wang
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.
Chunyan
C
Li
State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics (DICP), Chinese Academy of Sciences (CAS) , 457 Zhongshan Road, Dalian 116023, P. R. China.
Keli
K
Han
D000241:Adenosine; D043383:Adenosylhomocysteinase / Q000378:metabolism*; D007700:Kinetics; D008745:Methylation
D043383:Adenosylhomocysteinase; D000241:Adenosine
S-adenosylhomocysteine hydrolase; bioimaging; fluorogenic substrate; inhibitor screening; kinetic study