title: Gold nanoparticle supported phospholipid membranes as a biomimetic biosensor platform for phosphoinositide signaling detection.
abstract: Enzyme mediated phosphoinositide signaling plays important regulatory roles in diverse cellular processes and has close implication in human diseases. However, detection of phosphoinositide enzymes remains a challenge because of the difficulty in discriminating the phosphorylation patterns of phosphoinositide. Here we develop a novel enzyme-activated gold nanoparticles (AuNPs) assembly strategy as a homogeneous colorimetric biosensor for activity detection of phosphoinositide kinases and phosphatases. This strategy utilizes a biomimetic mechanism of phosphoinositide signaling, in which AuNP supported phospholipid membranes are constructed to mimic the cellular membrane substrate, and AuNPs modified with the pleckstrin homology (PH) domain of cytosolic proteins are designed for specific, multivalent recognition of phosphorylated phosphoinositides. This biomimetic strategy enables efficient enzymatic reactions of the substrate and highly selective detection of target enzyme. The biosensor is demonstrated for the detection of phosphoinositide 3-kinase (PI3K) and phosphatase with tensin homology (PTEN). The results revealed that it allows sensitive, rapid visual detection of the enzymes with pM detection limits and four-decade wide dynamic ranges, and is capable of detecting enzyme activities in complex cell lysate samples. This biosensor might provide a general biosensor platform for high-throughput detection of phosphoinositide enzymes with high sensitivity and selectivity in biomedical research and clinical diagnostics.
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Wen
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Si-Jia
authors.initials: SJ
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: tanglijuan@hnu.edu.cn.
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Tang
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China. Electronic address: jianhuijiang@hnu.edu.cn.
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Jiang
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D003124:Colorimetry; D057075:Enzyme Assays / Q000379:methods*; D006046:Gold / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D008567:Membranes, Artificial; D053768:Metal Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D051059:PTEN Phosphohydrolase / Q000032:analysis / Q000378:metabolism*; D019869:Phosphatidylinositol 3-Kinases / Q000032:analysis / Q000378:metabolism*; D010716:Phosphatidylinositols / Q000378:metabolism*; D010743:Phospholipids / Q000737:chemistry
chemical_list: D006046:Gold; D008567:Membranes, Artificial; D051059:PTEN Phosphohydrolase; D019869:Phosphatidylinositol 3-Kinases; D010716:Phosphatidylinositols; D010743:Phospholipids
keywords: Biosensor; Colorimetric detection; Gold nanoparticles; Phosphoinositide signaling; Phospholipid membranes