Measurement of kinase activation in single mammalian cells

被引：113

作者：

Meredith, GD ^{[1
]}

Sims, CE ^{[1
]}

Soughayer, JS ^{[1
]}

Allbritton, NL ^{[1
]}

机构：

[1] Univ Calif Irvine, Dept Physiol & Biophys, Irvine, CA 92697 USA

来源：

NATURE BIOTECHNOLOGY | 2000年 / 18卷 / 03期

关键词：

kinase; capillary zone electrophoresis; peptide; laser-micropipette;

D O I：

10.1038/73760

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：

We demonstrate a new method for the simultaneous measurement of the activation of key regulatory enzymes within single cells. To illustrate the capabilities of the technique, the activation of protein kinase C (PKC), protein kinase A (PKA), calcium-calmodulin activated kinase II (CamKII), and cdc2 protein kinase (cdc2K) was measured in response to both pharmacological or physiological stimuli. This assay strategy should be applicable to a broad range of intracellular enzymes, including phosphatases, proteases, nucleases, and other kinases.

引用

页码：309 / 312

页数：4

共 43 条

[1] SOURCE OF NUCLEAR CALCIUM SIGNALS [J].

ALLBRITTON, NL ;

OANCEA, E ;

KUHN, MA ;

MEYER, T .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (26) :12458-12462

[2] INOSITOL TRISPHOSPHATE AND CALCIUM SIGNALING [J].

BERRIDGE, MJ .

NATURE, 1993, 361 (6410) :315-325

[3]

BRIDGES AJ, 1997, ENCY CANC, P1314

[4] STUDIES WITH SYNTHETIC PEPTIDE-SUBSTRATES DERIVED FROM THE NEURONAL PROTEIN NEUROGRANIN REVEAL STRUCTURAL DETERMINANTS OF POTENCY AND SELECTIVITY FOR PROTEIN-KINASE-C [J].

CHEN, SJ ;

KLANN, E ;

GOWER, MC ;

POWELL, CM ;

SESSOMS, JS ;

SWEATT, JD .

BIOCHEMISTRY, 1993, 32 (04) :1032-1039

[5] THE STRUCTURE AND REGULATION OF PROTEIN PHOSPHATASES [J].

COHEN, P .

ANNUAL REVIEW OF BIOCHEMISTRY, 1989, 58 :453-508

[6] CAMP-DEPENDENT PROTEIN-KINASE, BUT NOT THE CGMP-DEPENDENT ENZYME, RAPIDLY PHOSPHORYLATES DELTA-CREB, AND A SYNTHETIC DELTA-CREB PEPTIDE [J].

COLBRAN, JL ;

ROACH, PJ ;

FIOL, CJ ;

DIXON, JE ;

ANDRISANI, OM ;

CORBIN, JD .

BIOCHEMISTRY AND CELL BIOLOGY-BIOCHIMIE ET BIOLOGIE CELLULAIRE, 1992, 70 (10-11) :1277-1282

[7] Calcium oscillations increase the efficiency and specificity of gene expression [J].

Dolmetsch, RE ;

Xu, KL ;

Lewis, RS .

NATURE, 1998, 392 (6679) :933-936

[8] Cell cycle checkpoints: Preventing an identity crisis [J].

Elledge, SJ .

SCIENCE, 1996, 274 (5293) :1664-1672

[9] SPONTANEOUS INJECTION IN MICROCOLUMN SEPARATIONS [J].

FISHMAN, HA ;

AMUDI, NM ;

LEE, TT ;

SCHELLER, RH ;

ZARE, RN .

ANALYTICAL CHEMISTRY, 1994, 66 (14) :2318-2329

[10] EXPRESSION OF A MULTIFUNCTIONAL CA-2+ CALMODULIN-DEPENDENT PROTEIN-KINASE AND MUTATIONAL ANALYSIS OF ITS AUTO-REGULATION [J].

HANSON, PI ;

KAPILOFF, MS ;

LOU, LL ;

ROSENFELD, MG ;

SCHULMAN, H .

NEURON, 1989, 3 (01) :59-70

← 1 2 3 4 5 →