title: Enzymatic immuno-assembly of gold nanoparticles for visualized activity screening of histone-modifying enzymes.
abstract: Activity screening of histone-modifying enzymes is of paramount importance for epigenetic research as well as clinical diagnostics and therapeutics. A novel biosensing strategy has been developed for sensitive and selective detection of histone-modifying enzymes as well as their inhibitors. This strategy relies on the antibody-mediated assembly of gold nanoparticles (AuNPs) decorated with substrate peptides that are subjected to enzymatic modifications by the histone-modifying enzymes. This design allows a visual and homogeneous assay of the enzyme activity using antibodies without any labels, which circumvents the requirements to prefunctionalize the antibody and affords improved assay simplicity and throughput. Additionally, the use of antibody-based recognition of modified peptides could offer improved specificity as compared with existing techniques based on the enzyme coupled assay. We have demonstrated this strategy using a histone methyltransferase acting on histone H3 (Lys 4) and a histone acetyltransferase acting on histone H3 (Lys 14). The results reveal that the absorption peak characteristic for AuNPs decreases dynamically with increasing activity of the enzymes with concomitant visualizable color attenuation, and subnanomolar detection limits are readily achieved for both enzymes. The developed strategy can thus offer a robust and convenient visualized platform for screening the enzyme activities and their inhibitors with high sensitivity and selectivity.
authors.affiliation: State Key Laboratory of Chemo/Bio-Sensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha, P R China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhen
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Tang
authors.forename: Haoxu
authors.initials: H
authors.lastname: Long
authors.forename: Jian-Hui
authors.initials: JH
authors.lastname: Jiang
mesh_terms: D015374:Biosensing Techniques; D004798:Enzymes / Q000032:analysis / Q000378:metabolism*; D006046:Gold / Q000737:chemistry*; D006657:Histones / Q000378:metabolism*; D007118:Immunoassay / Q000379:methods*; D057230:Limit of Detection; D053768:Metal Nanoparticles / Q000737:chemistry*
chemical_list: D004798:Enzymes; D006657:Histones; D006046:Gold