Mass Spectrometry-Based Structural Dissection of Fluorescent Proteins

被引：6

作者：

Alvarez, Luis A. ^{[2
]}

Merola, Fabienne ^{[2
]}

Erard, Marie ^{[2
]}

Rusconi, Filippo ^{[1
]}

机构：

[1] Museum Natl Hist Nat, Biophys Lab, CNRS, INSERM MNHN USM0503 UMR5153 U565, F-75231 Paris 05, France

[2] Univ Paris 11, Chim Phys Lab, CNRS, UMR 8000, F-91405 Orsay, France

来源：

BIOCHEMISTRY | 2009年 / 48卷 / 18期

关键词：

CHROMOPHORE; GFP; DECARBOXYLATION; OXIDATION; VARIANTS; CELLS; COLOR;

D O I：

10.1021/bi900327f

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Fluorescent proteins (FPs) are essential for live cell studies using fluorescence microscopy. To date, the molecular basis for FPs' irreversible photobleaching and the nature of the associated photoproducts are a matter of debate. Mass spectrometry, which should be an ideal technique for the structural dissection of FPs, cannot be harnessed efficiently due to their extreme resistance to trypsinolysis, due to the compactness of the barrel structure containing the chromopeptide. We devised a mild endoproteolysis procedure that affords a peptide mass fingerprint almost totally covering the sequence, thus allowing high-resolution mass spectrometric investigations of the protein structure.

引用

页码：3810 / 3812

页数：3

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