Rapid Detection of Protein Phosphatase Activity Using Zn(II)Coordinated Gold Nanosensors Based on His-Tagged Phosphopeptides

Title
Rapid Detection of Protein Phosphatase Activity Using Zn(II)Coordinated Gold Nanosensors Based on His-Tagged Phosphopeptides
Author
김태욱
Issue Date
2015-01
Publisher
AMER CHEMICAL SOC
Citation
ANALYTICAL CHEMISTRY, v. 87, NO 2, Page. 1257-1265
Abstract
We report a rapid colorimetric assay to detect protein phosphatase (PP) activity based on the controlled assembly and disassembly of gold nanoparticles (AuNPs) via Zn(II)-specific coordination in the presence of His(6)-tagged phosphopeptides. Among divalent metal ions including Ni(II), Cu(II), Co(II), Mg(II), Mn(II), and Zn(II), only Zn(II) triggered a strong association between phosphopeptides with hexahistidine at a single end and nitrilotriacetic acid (NTA)-modified AuNPs (21.3 nm in core diameter), leading to the self-assembly of AuNPs and consequently changes in color of the AuNP solution. In contrast, unphosphorylated peptides and His6-deficient phosphopeptides did not change the color of the AuNP solution. As a result, protein phosphatase 1 (PP1) activity and its inhibition were easily quantified with high sensitivity by determining the extinction ratio (E-520/E-700) of colloidal AuNPs. Most importantly, this method was capable of detecting protein phosphatase 2A (PP2A) activity in immunoprecipitated plant extracts. Because PPs play pivotal roles in mediating diverse signal transduction pathways as primary effectors of protein dephosphorylation, we anticipate that our method will be applied as a rapid format method to analyze the activities of various PPs and their inhibition.
URI
http://hdl.handle.net/20.500.11754/21241http://pubs.acs.org/doi/abs/10.1021/ac5039412
ISSN
0003-2700
DOI
10.1021/ac5039412
Appears in Collections:
COLLEGE OF NATURAL SCIENCES[S](자연과학대학) > LIFE SCIENCE(생명과학과) > Articles
Files in This Item:
There are no files associated with this item.
Export
RIS (EndNote)
XLS (Excel)
XML


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE