Labeling of fusion proteins with synthetic fluorophores in live cells

被引：337

作者：

Keppler, A ^{[1
]}

Pick, H ^{[1
]}

Arrivoli, C ^{[1
]}

Vogel, H ^{[1
]}

Johnsson, K ^{[1
]}

机构：

[1] Ecole Polytech Fed Lausanne, Inst Chem Sci & Engn, CH-1015 Lausanne, Switzerland

来源：

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA | 2004年 / 101卷 / 27期

关键词：

D O I：

10.1073/pnas.0401923101

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：

A general approach for the sequential labeling of fusion proteins of O-6-alkylguanine-DNA alkyltransferase (AGT) with different fluorophores in mammalian cells is presented. AGT fusion proteins with different localizations in the cell can be labeled specifically with different fluorophores, and the fluorescence labeling can be used for applications such as multicolor analysis of dynamic processes and fluorescence resonance energy transfer measurements. The facile access to a variety of different AGT substrates as well as the specificity of the labeling reaction should make the approach an important tool to study protein function in live cells.

引用

页码：9955 / 9959

页数：5

共 30 条

[1]

BALCH WE, 1986, J BIOL CHEM, V261, P4690

[2] IMMUNOELECTRON MICROSCOPIC STUDIES OF THE INTRACELLULAR-TRANSPORT OF THE MEMBRANE GLYCOPROTEIN (G) OF VESICULAR STOMATITIS-VIRUS IN INFECTED CHINESE-HAMSTER OVARY CELLS [J].

BERGMANN, JE ;

SINGER, SJ .

JOURNAL OF CELL BIOLOGY, 1983, 97 (06) :1777-1787

[3] Protein localization in living cells and tissues using FRET and FLIM [J].

Chen, Y ;

Mills, JD ;

Periasamy, A .

DIFFERENTIATION, 2003, 71 (9-10) :528-541

[4] SITE-SPECIFIC INCORPORATION OF BIOPHYSICAL PROBES INTO PROTEINS [J].

CORNISH, VW ;

BENSON, DR ;

ALTENBACH, CA ;

HIDEG, K ;

HUBBELL, WL ;

SCHULTZ, PG .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (08) :2910-2914

[5] QUALITY-CONTROL IN THE ENDOPLASMIC-RETICULUM - FOLDING AND MISFOLDING OF VESICULAR STOMATITIS-VIRUS G-PROTEIN IN CELLS AND INVITRO [J].

DESILVA, AM ;

BALCH, WE ;

HELENIUS, A .

JOURNAL OF CELL BIOLOGY, 1990, 111 (03) :857-866

[6] Fluorescence localization after photobleaching (FLAP): a new method for studying protein dynamics in living cells [J].

Dunn, GA ;

Dobbie, IM ;

Monypenny, J ;

Holt, MR ;

Zicha, D .

JOURNAL OF MICROSCOPY, 2002, 205 (01) :109-112

[7] INDUCIBILITY OF THE DNA-REPAIR GENE ENCODING O6-METHYLGUANINE-DNA METHYLTRANSFERASE IN MAMMALIAN-CELLS BY DNA-DAMAGING TREATMENTS [J].

FRITZ, G ;

TANO, K ;

MITRA, S ;

KAINA, B .

MOLECULAR AND CELLULAR BIOLOGY, 1991, 11 (09) :4660-4668

[8] Multicolor and electron microscopic imaging of connexin trafficking [J].

Gaietta, G ;

Deerinck, TJ ;

Adams, SR ;

Bouwer, J ;

Tour, O ;

Laird, DW ;

Sosinsky, GE ;

Tsien, RY ;

Ellisman, MH .

SCIENCE, 2002, 296 (5567) :503-507

[9] Induced protein dimerization in vivo through covalent labeling [J].

Gendreizig, S ;

Kindermann, M ;

Johnsson, K .

JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 2003, 125 (49) :14970-14971

[10] Specific covalent labeling of recombinant protein molecules inside live cells [J].

Griffin, BA ;

Adams, SR ;

Tsien, RY .

SCIENCE, 1998, 281 (5374) :269-272

← 1 2 3 →