Protein kinase assay on peptide-conjugated gold nanoparticles

被引：32

作者：

Kim, Young-Pil ^{[1
]}

Oh, Young-Hee ^{[1
]}

Kim, Hak-Sung ^{[1
]}

机构：

[1] Korea Adv Inst Sci & Technol, Dept Biol Sci, Taejon 305701, South Korea

来源：

BIOSENSORS & BIOELECTRONICS | 2008年 / 23卷 / 07期

关键词：

protein kinase; phosphorylation; gold nanoparticles; peptide; fluorescent assay;

D O I：

10.1016/j.bios.2007.10.001

中图分类号：

Q6 [生物物理学];

学科分类号：

071011 ;

摘要：

We demonstrate that protein kinase can be assayed with high sensitivity on peptide-conjugated gold nanoparticles (AuNPs). Phosphorylation of peptides on the AuNP-monolayers was detected by using an anti-phosphotyrosine antibody (alpha-pY) and Cy3-labeled secondary antibody (Cy3-alpha-mIgG) as a probing molecule. When compared to conventional self-assembled monolayers; (SAMs), spherical and three-dimensional geometry of AuNPs led to high surface density of peptide substrate and easy accessibility to enzyme, and consequently the resulting AuNP monolayers gave rise to improved detection sensitivity. Blocking of peptide-conjugated AuNPs with a poly(ethylene glycol) (PEG) also contributed to a higher signal-to-background ratio in kinase and its inhibition assays. The use of AuNPs as the platform surface will enable highly sensitive detection of protein kinases in a high-throughput manner. (C) 2007 Elsevier B.V. All rights reserved.

引用

页码：980 / 986

页数：7

共 19 条

[1] Gold nanoparticles: Assembly, supramolecular chemistry, quantum-size-related properties, and applications toward biology, catalysis, and nanotechnology [J].

Daniel, MC ;

Astruc, D .

CHEMICAL REVIEWS, 2004, 104 (01) :293-346

[2]

DAUCEY J, 2003, NAT REV DRUG DISCOV, V2, P296

[3] CONTROL OF THE SPECIFIC ADSORPTION OF PROTEINS ONTO GOLD SURFACES WITH POLY(L-LYSINE) MONOLAYERS [J].

FREY, BL ;

JORDAN, CE ;

KORNGUTH, S ;

CORN, RM .

ANALYTICAL CHEMISTRY, 1995, 67 (24) :4452-4457

[4] Two-dimensional arrays of colloidal gold particles: A flexible approach to macroscopic metal surfaces [J].

Grabar, KC ;

Allison, KJ ;

Baker, BE ;

Bright, RM ;

Brown, KR ;

Freeman, RG ;

Fox, AP ;

Keating, CD ;

Musick, MD ;

Natan, MJ .

LANGMUIR, 1996, 12 (10) :2353-2361

[5] Kinetic and equilibrium binding analysis of protein-ligand interactions at poly(amidoamine) dendrimer monolayers [J].

Hong, MY ;

Lee, D ;

Kim, HS .

ANALYTICAL CHEMISTRY, 2005, 77 (22) :7326-7334

[6] Peptide chips for the quantitative evaluation of protein kinase activity [J].

Houseman, BT ;

Huh, JH ;

Kron, SJ ;

Mrksich, M .

NATURE BIOTECHNOLOGY, 2002, 20 (03) :270-274

[7] Gold nanoparticle-enhanced secondary ion mass spectrometry imaging of peptides on self-assembled monolayers [J].

Kim, YP ;

Oh, E ;

Hong, MY ;

Lee, D ;

Han, MK ;

Shon, HK ;

Moon, DW ;

Kim, HS ;

Lee, TG .

ANALYTICAL CHEMISTRY, 2006, 78 (06) :1913-1920

[8] Printing small molecules as microarrays and detecting protein-ligand interactions en masse [J].

MacBeath, G ;

Koehler, AN ;

Schreiber, SL .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1999, 121 (34) :7967-7968

[9] CRYSTALLOGRAPHIC REFINEMENT AND ATOMIC MODELS OF THE INTACT IMMUNOGLOBULIN MOLECULE KOL AND ITS ANTIGEN-BINDING FRAGMENT AT 3.0-A AND 1.9-A RESOLUTION [J].

MARQUART, M ;

DEISENHOFER, J ;

HUBER, R ;

PALM, W .

JOURNAL OF MOLECULAR BIOLOGY, 1980, 141 (04) :369-391

[10] Quantitative methods for the analysis of protein phosphorylation in drug development [J].

Olive, DM .

EXPERT REVIEW OF PROTEOMICS, 2004, 1 (03) :327-341

← 1 2 →