title: Real time monitoring of on-chip coenzyme regeneration with SPR and DPI.
abstract: We report in this work real time characterization of enzyme-coenzyme binding by using surface plasmon resonance (SPR) and dual polarization interferometry (DPI) analyses. Results showed that diaphorase (DP) and lactate dehydrogenases (LDH) had distinct binding selectivity and preference over reduced and oxidized states of coenzyme NAD(H). On the basis of that, DP and LDH were chosen as indicator enzymes to distinguish the specific state of surface-bound NAD(H). The transformation between NADH and NAD(+) during enzyme-catalyzed redox reactions was therefore transduced into variation in interaction signals as indicated via the binding status of the indicator enzymes as detected with both SPR and DPI. This real time molecule-specific detection strategy revealed quick and direct reflection of the state and reactivity of the coenzyme, promising a unique way of precise molecular interaction analysis.
authors: National Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100190, China.
authors: Xiaoyi
authors: X
authors: Feng
authors: Fei
authors: F
authors: Gao
authors: Peiyong
authors: P
authors: Qin
authors: Guanghui
authors: G
authors: Ma
authors: Zhiguo
authors: Z
authors: Su
authors: Jia
authors: J
authors: Ge
authors: Ping
authors: P
authors: Wang
authors: Songping
authors: S
authors: Zhang
mesh_terms: D055162:Biocatalysis; D003911:Dextrans / Q000737:chemistry; D007368:Interferometry; D007770:L-Lactate Dehydrogenase / Q000378:metabolism*; D009243:NAD / Q000378:metabolism*; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000378:metabolism; D011485:Protein Binding; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
chemical_list: D003911:Dextrans; D009243:NAD; D010088:Oxidoreductases; D007770:L-Lactate Dehydrogenase