An improved smaller biotin ligase for BioID proximity labeling

被引：542

作者：

Kim, Dae In ^{[1
]}

Jensen, Samuel C. ^{[1
]}

Noble, Kyle A. ^{[2
]}

Birendra, K. C. ^{[1
]}

Roux, Kenneth H. ^{[2
]}

Motamedchaboki, Khatereh ^{[3
]}

Roux, Kyle J. ^{[1
,4
]}

机构：

[1] Sanford Childrens Hlth Res Ctr, Sanford Res, Sioux Falls, SD 57104 USA

[2] Florida State Univ, Dept Biol Sci, B-157, Tallahassee, FL 32306 USA

[3] Sanford Burnham Med Res Inst, Sanford Burnham Prote Facil, La Jolla, CA 92037 USA

[4] Univ S Dakota, Sanford Sch Med, Dept Pediat, Sioux Falls, SD 57105 USA

来源：

MOLECULAR BIOLOGY OF THE CELL | 2016年 / 27卷 / 08期

基金：

美国国家卫生研究院;

关键词：

NUCLEAR-PORE COMPLEX; PROMISCUOUS PROTEIN BIOTINYLATION; MESSENGER-RNA; INTERACTING PARTNERS; PROTEOMIC ANALYSIS; E-CADHERIN; IDENTIFICATION; MEMBRANE; PATHWAY; CELL;

D O I：

10.1091/mbc.E15-12-0844

中图分类号：

Q2 [细胞生物学];

学科分类号：

071009 ; 090102 ;

摘要：

The BioID method uses a promiscuous biotin ligase to detect protein-protein associations as well as proximate proteins in living cells. Here we report improvements to the BioID method centered on BioID2, a substantially smaller promiscuous biotin ligase. BioID2 enables more-selective targeting of fusion proteins, requires less biotin supplementation, and exhibits enhanced labeling of proximate proteins. Thus BioID2 improves the efficiency of screening for protein-protein associations. We also demonstrate that the biotinylation range of BioID2 can be considerably modulated using flexible linkers, thus enabling application-specific adjustment of the biotin-labeling radius.

引用

页码：1188 / 1196

页数：9

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