Rapid parallel flow cytometry assays of active GTPases using effector beads

被引：15

作者：

Buranda, Tione ^{[1
,2
,3
]}

BasuRay, Soumik ^{[1
]}

Swanson, Scarlett ^{[1
]}

Agola, Jacob ^{[1
]}

Bondu, Virginie ^{[1
]}

Wandinger-Ness, Angela ^{[1
,2
]}

机构：

[1] Univ New Mexico, Sch Med, Dept Pathol, Albuquerque, NM 87131 USA

[2] Univ New Mexico, Sch Med, Ctr Canc, Albuquerque, NM 87131 USA

[3] Univ New Mexico, Sch Med, Ctr Infect Dis & Immun, Albuquerque, NM 87131 USA

来源：

ANALYTICAL BIOCHEMISTRY | 2013年 / 442卷 / 02期

基金：

美国国家卫生研究院; 美国国家科学基金会;

关键词：

Rho; Rac; Cdc42; Rab GTPases; Hantavirus; lntegrins; DECAY-ACCELERATING FACTOR; BCL-2 FAMILY PROTEINS; RHO-GTPASES; NUCLEOTIDE-BINDING; GROWTH-FACTOR; CELL; ACTIVATION; KINASE; RAS; PATHWAY;

D O I：

10.1016/j.ab.2013.07.039

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

We describe a rapid assay for measuring the cellular activity of small guanine triphosphatases (GTPases) in response to a specific stimulus. Effector-functionalized beads are used to quantify in parallel multiple GTP-bound GTPases in the same cell lysate by flow cytometry. In a biologically relevant example, five different Ras family GTPases are shown for the first time to be involved in a concerted signaling cascade downstream of receptor ligation by Sin Nombre hantavirus. (C) 2013 Elsevier Inc. All rights reserved.

引用

页码：149 / 157

页数：9

共 47 条

[1] A Competitive Nucleotide Binding Inhibitor: In Vitro Characterization of Rab7 GTPase Inhibition [J].

Agola, Jacob O. ;

Hong, Lin ;

Surviladze, Zurab ;

Ursu, Oleg ;

Waller, Anna ;

Strouse, J. Jacob ;

Simpson, Denise S. ;

Schroeder, Chad E. ;

Oprea, Tudor I. ;

Golden, Jennifer E. ;

Aube, Jeffrey ;

Buranda, Tione ;

Sklar, Larry A. ;

Wandinger-Ness, Angela .

ACS CHEMICAL BIOLOGY, 2012, 7 (06) :1095-1108

[2] The Rac1-and RhoG-specific GEF domain of Trio targets filamin to remodel cytoskeletal actin [J].

Bellanger, JM ;

Astier, C ;

Sardet, C ;

Ohta, Y ;

Stossel, TP ;

Debant, A .

NATURE CELL BIOLOGY, 2000, 2 (12) :888-892

[3] Intramuscular inoculation of Sin Nombre hantavirus cDNAs induces cellular and humoral immune responses in BALB c mice [J].

Bharadwaj, M ;

Lyons, CR ;

Wortman, IA ;

Hjelle, B .

VACCINE, 1999, 17 (22) :2836-2843

[4] THE SMALL GTPASE RAB5 FUNCTIONS AS A REGULATORY FACTOR IN THE EARLY ENDOCYTIC PATHWAY [J].

BUCCI, C ;

PARTON, RG ;

MATHER, IH ;

STUNNENBERG, H ;

SIMONS, K ;

HOFLACK, B ;

ZERIAL, M .

CELL, 1992, 70 (05) :715-728

[5] Recognition of decay accelerating factor and αvβ3 by inactivated hantaviruses: Toward the development of high-throughput screening flow cytometry assays [J].

Buranda, Tione ;

Wu, Yang ;

Perez, Dominique ;

Jett, Stephen D. ;

BonduHawkins, Virginie ;

Ye, Chunyan ;

Edwards, Bruce ;

Hall, Pamela ;

Larson, Richard S. ;

Lopez, Gabriel P. ;

Sklar, Larry A. ;

Hjelle, Brian .

ANALYTICAL BIOCHEMISTRY, 2010, 402 (02) :151-160

[6] Rho and Rac take center stage [J].

Burridge, K ;

Wennerberg, K .

CELL, 2004, 116 (02) :167-179

[7] Front and back by Rho and Rac [J].

Burridge, Keith ;

Doughman, Renee .

NATURE CELL BIOLOGY, 2006, 8 (08) :781-782

[8] REGULATION OF SMALL GTPases BY GEFs, GAPs, AND GDIs [J].

Cherfils, Jacqueline ;

Zeghouf, Mahel .

PHYSIOLOGICAL REVIEWS, 2013, 93 (01) :269-309

[9] FRET detection of cellular α4-integrin conformational activation [J].

Chigaev, A ;

Buranda, T ;

Dwyer, DC ;

Prossnitz, ER ;

Sklar, LA .

BIOPHYSICAL JOURNAL, 2003, 85 (06) :3951-3962

[10] Expression, assay, and functional properties of RILP [J].

Colucci, AMR ;

Spinosa, MR ;

Bucci, C .

GTPASES REGULATING MEMBRANE TARGETING AND FUSION, 2005, 403 :664-+

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