Multiphoton excitation-evoked chromophore-assisted laser inactivation using green fluorescent protein

Takuji Tanabe; Masahito Oyamada; Katsumasa Fujita; Ping Dai; Hideo Tanaka; Tetsuro Takamatsu

doi:10.1038/nmeth770

Multiphoton excitation-evoked chromophore-assisted laser inactivation using green fluorescent protein

Nat Methods. 2005 Jul;2(7):503-5. doi: 10.1038/nmeth770.

Authors

Takuji Tanabe¹, Masahito Oyamada, Katsumasa Fujita, Ping Dai, Hideo Tanaka, Tetsuro Takamatsu

Affiliation

¹ Department of Pathology and Cell Regulation, Kyoto Prefectural University of Medicine, 465 Kajii-cho Hirokoji Kawaramachi, Kamigyo-ku, Kyoto 602-8566, Japan.

PMID: 15973419
DOI: 10.1038/nmeth770

Abstract

Noninvasive, straightforward methods to inactivate selected proteins in living cells with high spatiotemporal resolution are needed. Chromophore-assisted laser inactivation (CALI) can be used to photochemically inactivate proteins, but it has several drawbacks, such as procedural complexity and nonspecific photodamage. Here we show that by application of multiphoton excitation to CALI, enhanced green fluorescent protein (EGFP) is an effective chromophore for inactivation of a protein's function without nonspecific photodamage in living mammalian cells.

Publication types

Evaluation Study
Research Support, Non-U.S. Gov't

MeSH terms

Connexin 43 / genetics
Connexin 43 / metabolism*
Fluorescent Dyes
Gene Targeting / methods*
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism*
HeLa Cells
Humans
Lasers*
Microscopy, Confocal / methods*
Microscopy, Fluorescence, Multiphoton / methods*
Recombinant Fusion Proteins / genetics
Recombinant Fusion Proteins / metabolism
Staining and Labeling / methods

Substances

Connexin 43
Fluorescent Dyes
Recombinant Fusion Proteins
enhanced green fluorescent protein
Green Fluorescent Proteins