title: Label-free fluorescent enzymatic assay of citrate synthase by CoA-Au(I) co-ordination polymer and its application in a multi-enzyme logic gate cascade.
abstract: Citrate synthase (CS) is one of the key metabolic enzymes in the Krebs tricarboxylic acid (TCA) cycle. It regulates energy generation in mitochondrial respiration by catalysing the reaction between oxaloacetic acid (OAA) and acetyl coenzyme A (Ac-CoA) to generate citrate and coenzyme A (CoA). CS has been shown to be a biomarker of neurological diseases and various kinds of cancers. Here, a label-free fluorescent assay has been developed for homogeneously detecting CS and its inhibitor based on the in situ generation of CoA-Au(I) co-ordination polymer (CP) and the fluorescence signal-on by SYBR Green II-stained CoA-Au(I) CP. Because of the unique property of the CoA-Au(I) CP, this CS activity assay method could achieve excellent selectivity and sensitivity, with a linear range from 0.0033 U/μL to 0.264 U/μL and a limit of detection to be 0.00165 U/μL. Meanwhile, this assay method has advantages of being facile and cost effective with quick detection. Moreover, based on this method, a biomimetic logic system was established by rationally exploiting the cascade enzymatic interactions in TCA cycle for chemical information processing. In the TCA cycle-derived logic system, an AND-AND-AND-cascaded gate was rigorously operated step by step in one pot, and is outputted by a label-free fluorescent signal with visualized readout.
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Yong
authors: Y
authors: Li
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Huixia
authors: H
authors: Wang
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Futao
authors: F
authors: Dai
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Pei
authors: P
authors: Li
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Xin
authors: X
authors: Jin
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Yan
authors: Y
authors: Huang
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China. Electronic address: niezhou.hnu@gmail.com.
authors: Zhou
authors: Z
authors: Nie
authors: State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, China.
authors: Shouzhuo
authors: S
authors: Yao
mesh_terms: D000105:Acetyl Coenzyme A / Q000737:chemistry*; D002950:Citrate (si)-Synthase / Q000032:analysis* / Q000737:chemistry; D004789:Enzyme Activation; D005456:Fluorescent Dyes / Q000138:chemical synthesis; D009097:Multienzyme Complexes / Q000032:analysis* / Q000737:chemistry*; D062907:Oxaloacetic Acid / Q000737:chemistry*; D012815:Signal Processing, Computer-Assisted; D013050:Spectrometry, Fluorescence / Q000379:methods*; D013194:Staining and Labeling
chemical_list: D005456:Fluorescent Dyes; D009097:Multienzyme Complexes; D062907:Oxaloacetic Acid; D000105:Acetyl Coenzyme A; D002950:Citrate (si)-Synthase
keywords: Citrate synthase activity assay; Label-free fluorometry; Logic gate cascade; Thiol–Au(I) co-ordination polymer