A Redox-Sensitive Luciferase Assay for Determining the Localization and Topology of Endoplasmic Reticulum Proteins

被引：9

作者：

Li, Hai-Yin ^{[1
]}

Zheng, Xue-Ming ^{[1
]}

Che, Mei-Xia ^{[1
]}

Hu, Hong-Yu ^{[1
]}

机构：

[1] Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai, Peoples R China

来源：

PLOS ONE | 2012年 / 7卷 / 04期

基金：

中国国家自然科学基金;

关键词：

GREEN FLUORESCENT PROTEIN; GAUSSIA LUCIFERASE; SECRETORY PATHWAY; QUALITY-CONTROL; DEGRADATION; STRESS; DOMAIN;

D O I：

10.1371/journal.pone.0035628

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

Correct localization and transmembrane topology are crucial for the proteins residing and functioning in the endoplasmic reticulum (ER). We have developed a rapid and convenient assay, based on the redox-sensitive luciferase from Gaussia princeps (Gluc) and green fluorescence protein (GFP), to determine the localization or topology of ER proteins. Using the tandem Gluc-GFP reporter fused to different positions of a target protein, we successfully characterized the topologies of two ER transmembrane proteins Herp and HRD1 that are involved in the ER quality control system. This assay method may also be applicable to the proteins in secretory pathway, plasma membrane, and other compartments of cells.

引用

页数：7

共 22 条

[1] A Highly Sensitive Assay for Monitoring the Secretory Pathway and ER Stress [J].

Badr, Christian E. ;

Hewett, Jeffrey W. ;

Breakefield, Xandra O. ;

Tannous, Bakhos A. .

PLOS ONE, 2007, 2 (06)

[2] Structure of Monomeric Yeast and Mammalian Sec61 Complexes Interacting with the Translating Ribosome [J].

Becker, Thomas ;

Bhushan, Shashi ;

Jarasch, Alexander ;

Armache, Jean-Paul ;

Funes, Soledad ;

Jossinet, Fabrice ;

Gumbart, James ;

Mielke, Thorsten ;

Berninghausen, Otto ;

Schulten, Klaus ;

Westhof, Eric ;

Gilmore, Reid ;

Mandon, Elisabet C. ;

Beckmann, Roland .

SCIENCE, 2009, 326 (5958) :1369-1373

[3] Stringent requirement for HRD1, SEL1L, and OS-9/XTP3-B for disposal of ERAD-LS substrates [J].

Bernasconi, Riccardo ;

Galli, Carmela ;

Calanca, Verena ;

Nakajima, Toshihiro ;

Molinari, Maurizio .

JOURNAL OF CELL BIOLOGY, 2010, 188 (02) :223-235

[4] Non-invasive topology analysis of membrane proteins in the secretory pathway [J].

Brach, Thorsten ;

Soyk, Sebastian ;

Mueller, Christopher ;

Hinz, Giselbert ;

Hell, Ruediger ;

Brandizzi, Federica ;

Meyer, Andreas J. .

PLANT JOURNAL, 2009, 57 (03) :534-541

[5] Protein Quality Control in the Cytosol and the Endoplasmic Reticulum: Brothers in Arms [J].

Buchberger, Alexander ;

Bukau, Bernd ;

Sommer, Thomas .

MOLECULAR CELL, 2010, 40 (02) :238-252

[6] Diffusion of green fluorescent protein in the aqueous-phase lumen of endoplasmic reticulum [J].

Dayel, MJ ;

Hom, EFY ;

Verkman, AS .

BIOPHYSICAL JOURNAL, 1999, 76 (05) :2843-2851

[7] Membrane topology and function of Der3/Hrd1p as a ubiquitin-protein ligase (E3) involved in endoplasmic reticulum degradation [J].

Deak, PM ;

Wolf, DH .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2001, 276 (14) :10663-10669

[8] Quality control in the endoplasmic reticulum [J].

Ellgaard, L ;

Helenius, A .

NATURE REVIEWS MOLECULAR CELL BIOLOGY, 2003, 4 (03) :181-191

[9] Identification of two catalytic domains in a luciferase secreted by the copepod Gaussia princeps [J].

Inouye, Satoshi ;

Sahara, Yuiko .

BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2008, 365 (01) :96-101

[10] Herp, a new ubiquitin-like membrane protein induced by endoplasmic reticulum stress [J].

Kokame, K ;

Agarwala, KL ;

Kato, H ;

Miyata, T .

JOURNAL OF BIOLOGICAL CHEMISTRY, 2000, 275 (42) :32846-32853

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