New Approach for Local Structure Analysis of the Tyrosine Domain in Proteins by Using a Site-Specific and Polarity-Sensitive Fluorescent Probe

doi:10.1002/cbic.200900003

CORC > 化学研究所 > 中国科学院化学研究所

	New Approach for Local Structure Analysis of the Tyrosine Domain in Proteins by Using a Site-Specific and Polarity-Sensitive Fluorescent Probe
	Chen, Suming; Li, Xiaohua; Ma, Huimin
刊名	CHEMBIOCHEM
	2009-05-04
卷号	10 期号:7 页码:1200-1207
关键词	Analytical Methods Fluorescent Probes Local Polarity Detection Transition Metals Tyrosine
ISSN号	1439-4227
DOI	10.1002/cbic.200900003
英文摘要	The design and synthesis of a novel long-wavelength polarity-sensitive fluorescence probe, 6-[9-(diethylamino)-5-oxo-5H-benzo[alpha]phenoxazin-2-yloxy]hex-2-enyl acetate, for the selective modification of tyrosine residues with the goal of providing local information on tyrosine domains in proteins, is reported. This probe comprises a polarity-sensitive Nile red fluorophore and an active pi-allyl group that can form pi-allyipalladium complexes and react selectively with tyrosine residues. The probe has the following features: 1) it has a long-wavelength emission of > 550nm, thanks to which interference from short-wavelength fluorescence from common biological matrixes can be avoided; 2) the maximum emission wavelength is sensitive only to polarity and not to pH or temperature; this allows the accurate determination of local polarity; and 3) it is a neutral, uncharged molecule, and does not disturb,the over-all charge of the labelled protein. With this probe the polarity and conformation changes of the Tyr108 domain in native and in acid- and heat-denatured bovine Cu/Zn superoxide dismutase were detected for the first time. It was found that the polarity of the Tyr108 domain hardly alters on acid denaturation between pH 4 and 9. However, heat denaturation caused the Tyr108 domain to be more hydrophobic, and was accompanied by an irreversible aggregation of the protein. In addition, the probe-binding experiments revealed that the surface of the protein becomes more hydrophobic after thermal denaturation; this can be ascribed to the formation of the more hydrophobic aggregates. This strategy might provide a general approach for studying the local environment changes of tyrosine domains in proteins under acid or heat denaturation conditions.
语种	英语
出版者	WILEY-V C H VERLAG GMBH
WOS记录号	WOS:000266382900011
内容类型	期刊论文
源URL	[http://ir.iccas.ac.cn/handle/121111/67513]
专题	中国科学院化学研究所
通讯作者	Ma, Huimin
作者单位	Chinese Acad Sci, Beijing Natl Lab Mol Sci, Inst Chem, Beijing 100190, Peoples R China
推荐引用方式 GB/T 7714	Chen, Suming,Li, Xiaohua,Ma, Huimin. New Approach for Local Structure Analysis of the Tyrosine Domain in Proteins by Using a Site-Specific and Polarity-Sensitive Fluorescent Probe[J]. CHEMBIOCHEM,2009,10(7):1200-1207.
APA	Chen, Suming,Li, Xiaohua,&Ma, Huimin.(2009).New Approach for Local Structure Analysis of the Tyrosine Domain in Proteins by Using a Site-Specific and Polarity-Sensitive Fluorescent Probe.CHEMBIOCHEM,10(7),1200-1207.
MLA	Chen, Suming,et al."New Approach for Local Structure Analysis of the Tyrosine Domain in Proteins by Using a Site-Specific and Polarity-Sensitive Fluorescent Probe".CHEMBIOCHEM 10.7(2009):1200-1207.