Characterization of a rabbit polyclonal antibody against threonine-AMPylation

被引：29

作者：

Hao, Yi-Heng ^{[1
]}

Chuang, Trinette ^{[2
]}

Ball, Haydn L. ^{[3
]}

Luong, Phi ^{[1
]}

Li, Yan ^{[3
]}

Flores-Saaib, Ruben D. ^{[2
]}

Orth, Kim ^{[1
]}

机构：

[1] Univ Texas SW Med Ctr Dallas, Dept Mol Biol, Dallas, TX 75390 USA

[2] EMD Millipore, Temecula, CA 92590 USA

[3] Univ Texas SW Med Ctr Dallas, Dept Internal Med, Prot Chem Technol Ctr, Dallas, TX 75390 USA

来源：

JOURNAL OF BIOTECHNOLOGY | 2011年 / 151卷 / 03期

关键词：

AMP; AMPylation; Antibody; Western blot; Immunoprecipitation; FIC DOMAIN; PROTEIN; ADENYLYLATION;

D O I：

10.1016/j.jbiotec.2010.12.013

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

An antibody against the posttranslational modification AMPylation was produced using a peptide corresponding to human Rac1 switch I region with AMPylated threonine-35 residue as an antigen. The resulting rabbit antiserum was tested for its abilities to recognize AMPylated proteins by western blot and immunoprecipitation. The antiserum is highly specific for threonine-AMPylated proteins and weakly recognizes tyrosine-AMPylated proteins. Depletion of serum with modified protein abolished its activity against tyrosine-AMPylated proteins. The antiserum also recognized native proteins with modification in an immunoprecipitation experiment. Interactions of the antiserum could be inhibited by competition with AMP but not with GMP or UMP. This antiserum had potential utility for the identification of unknown AMPylated proteins. (C) 2010 Elsevier B.V. All rights reserved.

引用

页码：251 / 254

页数：4

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