A photoactivatable GFP for selective photolabeling of proteins and cells

被引：1186

作者：

Patterson, GH ^{[1
]}

Lippincott-Schwartz, J ^{[1
]}

机构：

[1] NICHHD, Cell Biol & Metab Branch, NIH, Bethesda, MD 20892 USA

来源：

SCIENCE | 2002年 / 297卷 / 5588期

关键词：

D O I：

10.1126/science.1074952

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：

We report a photoactivatable variant of the Aequorea victoria green fluorescent protein (GFP) that, after intense irradiation with 413-nanometer light, increases fluorescence 100 times when excited by 488-nanometer light and remains stable for days under aerobic conditions. These characteristics offer a new tool for exploring intracellular protein dynamics by tracking photoactivated molecules that are the only visible GFPs in the cell. Here, we use the photoactivatable GFP both as a free protein to measure protein diffusion across the nuclear envelope and as a chimera with a lysosomal membrane protein to demonstrate rapid interlysosomal membrane exchange.

引用

页码：1873 / 1877

页数：5

共 25 条

[1] Structural basis for dual excitation and photoisomerization of the Aequorea victoria green fluorescent protein [J].

Brejc, K ;

Sixma, TK ;

Kitts, PA ;

Kain, SR ;

Tsien, RY ;

Ormo, M ;

Remington, SJ .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1997, 94 (06) :2306-2311

[2] GREEN FLUORESCENT PROTEIN AS A MARKER FOR GENE-EXPRESSION [J].

CHALFIE, M ;

TU, Y ;

EUSKIRCHEN, G ;

WARD, WW ;

PRASHER, DC .

SCIENCE, 1994, 263 (5148) :802-805

[3] ANIMAL-CELL LYSOSOMES RAPIDLY EXCHANGE MEMBRANE-PROTEINS [J].

DENG, YP ;

STORRIE, B .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (11) :3860-3864

[4] GREEN-FLUORESCENT PROTEIN MUTANTS WITH ALTERED FLUORESCENCE EXCITATION-SPECTRA [J].

EHRIG, T ;

OKANE, DJ ;

PRENDERGAST, FG .

FEBS LETTERS, 1995, 367 (02) :163-166

[5] Photoactivation turns green fluorescent protein red [J].

Elowitz, MB ;

Surette, MG ;

Wolf, PE ;

Stock, J ;

Leibler, S .

CURRENT BIOLOGY, 1997, 7 (10) :809-812

[6] CHINESE-HAMSTER OVARY CELL LYSOSOMES RAPIDLY EXCHANGE CONTENTS [J].

FERRIS, AL ;

BROWN, JC ;

PARK, RD ;

STORRIE, B .

JOURNAL OF CELL BIOLOGY, 1987, 105 (06) :2703-2712

[7] Dynamics of fluorescence fluctuations in green fluorescent protein observed by fluorescence correlation spectroscopy [J].

Haupts, U ;

Maiti, S ;

Schwille, P ;

Webb, WW .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1998, 95 (23) :13573-13578

[8] WAVELENGTH MUTATIONS AND POSTTRANSLATIONAL AUTOXIDATION OF GREEN FLUORESCENT PROTEIN [J].

HEIM, R ;

PRASHER, DC ;

TSIEN, RY .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1994, 91 (26) :12501-12504

[9] DERIVED PROTEIN-SEQUENCE, OLIGOSACCHARIDES, AND MEMBRANE INSERTION OF THE 120-KDA LYSOSOMAL MEMBRANE GLYCOPROTEIN (LGP120) - IDENTIFICATION OF A HIGHLY CONSERVED FAMILY OF LYSOSOMAL MEMBRANE-GLYCOPROTEINS [J].

HOWE, CL ;

GRANGER, BL ;

HULL, M ;

GREEN, SA ;

GABEL, CA ;

HELENIUS, A ;

MELLMAN, I .

PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1988, 85 (20) :7577-7581

[10] THE BIOGENESIS OF LYSOSOMES [J].

KORNFELD, S ;

MELLMAN, I .

ANNUAL REVIEW OF CELL BIOLOGY, 1989, 5 :483-525

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