绿色荧光蛋白的单分子共聚焦显微镜

G Jung, J Wiehler, W Göhde, J Tittel, Th Basché, B Steipe, C Bräuchle
{"title":"绿色荧光蛋白的单分子共聚焦显微镜","authors":"G Jung,&nbsp;J Wiehler,&nbsp;W Göhde,&nbsp;J Tittel,&nbsp;Th Basché,&nbsp;B Steipe,&nbsp;C Bräuchle","doi":"10.1002/1361-6374(199803)6:1<54::AID-BIO7>3.0.CO;2-8","DOIUrl":null,"url":null,"abstract":"<p>Single molecule detection has been extended into life sciences by use of strongly fluorescent labels. The green fluorescent protein (GFP) as a self-fluorescent biomolecule has attracted considerable attention. Here, single molecules of the GFP-mutant Glu222Gln are immobilized in a polyvinylalcohol matrix and detected by confocal fluorescence microscopy. Although this mutant stabilizes one of both conformers of the wild-type GFP, the investigation of its fluorescence dynamics reveals strong signal fluctuations. This fluorescence behaviour is—at least partly—caused by reversible photochemical changes of the protein framework, that can relax into the fluorescent state on different timescales. Thus, this protein appears particularly appropriate for studying the microheterogeneity of the macromolecule GFP on a single molecule level.</p>","PeriodicalId":100176,"journal":{"name":"Bioimaging","volume":"6 1","pages":"54-61"},"PeriodicalIF":0.0000,"publicationDate":"2001-05-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/1361-6374(199803)6:1<54::AID-BIO7>3.0.CO;2-8","citationCount":"43","resultStr":"{\"title\":\"Confocal microscopy of single molecules of the green fluorescent protein\",\"authors\":\"G Jung,&nbsp;J Wiehler,&nbsp;W Göhde,&nbsp;J Tittel,&nbsp;Th Basché,&nbsp;B Steipe,&nbsp;C Bräuchle\",\"doi\":\"10.1002/1361-6374(199803)6:1<54::AID-BIO7>3.0.CO;2-8\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Single molecule detection has been extended into life sciences by use of strongly fluorescent labels. The green fluorescent protein (GFP) as a self-fluorescent biomolecule has attracted considerable attention. Here, single molecules of the GFP-mutant Glu222Gln are immobilized in a polyvinylalcohol matrix and detected by confocal fluorescence microscopy. Although this mutant stabilizes one of both conformers of the wild-type GFP, the investigation of its fluorescence dynamics reveals strong signal fluctuations. This fluorescence behaviour is—at least partly—caused by reversible photochemical changes of the protein framework, that can relax into the fluorescent state on different timescales. Thus, this protein appears particularly appropriate for studying the microheterogeneity of the macromolecule GFP on a single molecule level.</p>\",\"PeriodicalId\":100176,\"journal\":{\"name\":\"Bioimaging\",\"volume\":\"6 1\",\"pages\":\"54-61\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2001-05-11\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/1361-6374(199803)6:1<54::AID-BIO7>3.0.CO;2-8\",\"citationCount\":\"43\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Bioimaging\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/1361-6374%28199803%296%3A1%3C54%3A%3AAID-BIO7%3E3.0.CO%3B2-8\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioimaging","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/1361-6374%28199803%296%3A1%3C54%3A%3AAID-BIO7%3E3.0.CO%3B2-8","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 43

摘要

单分子检测已通过使用强荧光标记扩展到生命科学。绿色荧光蛋白(GFP)作为一种具有自荧光特性的生物分子受到了广泛的关注。在这里,gfp突变体Glu222Gln的单分子固定在聚乙烯醇基质中,并通过共聚焦荧光显微镜检测。虽然该突变体稳定了野生型GFP的两种构象之一,但对其荧光动力学的研究显示出强烈的信号波动。这种荧光行为——至少部分是由蛋白质框架的可逆光化学变化引起的,这种变化可以在不同的时间尺度上放松到荧光状态。因此,该蛋白似乎特别适合在单分子水平上研究大分子GFP的微观异质性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Confocal microscopy of single molecules of the green fluorescent protein

Single molecule detection has been extended into life sciences by use of strongly fluorescent labels. The green fluorescent protein (GFP) as a self-fluorescent biomolecule has attracted considerable attention. Here, single molecules of the GFP-mutant Glu222Gln are immobilized in a polyvinylalcohol matrix and detected by confocal fluorescence microscopy. Although this mutant stabilizes one of both conformers of the wild-type GFP, the investigation of its fluorescence dynamics reveals strong signal fluctuations. This fluorescence behaviour is—at least partly—caused by reversible photochemical changes of the protein framework, that can relax into the fluorescent state on different timescales. Thus, this protein appears particularly appropriate for studying the microheterogeneity of the macromolecule GFP on a single molecule level.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Magnetic Particle Imaging Magnetic Resonance Imaging Quantitative evaluation of light microscopes based on image processing techniques Confocal microscopy of single molecules of the green fluorescent protein Heavy metal contrast enhancement for the selective detection of gold particles in electron microscopical sections using electron spectroscopic imaging
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1