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相关概念视频

Reporter Genes02:11

Reporter Genes

Reporter genes are a type of protein-coding gene that are often tagged to a gene of interest. Once inside a target cell, reporter genes usually produce visually identifiable characteristics like fluorescence and luminescence when expressed along with the gene of interest. Thus, reporter genes “report” the presence or absence of genes of interest in an organism, determine the gene expression pattern, or track the physical location of a DNA segment or protein in the cell.
Commonly used reporter...
G-Protein Gated Ion Channels01:21

G-Protein Gated Ion Channels

GPCRs are primarily responsible for our sense of smell, taste, and vision.  The binding of a sensory stimulus activates GPCR to stimulate effector proteins, many of which are ion channels in the sensory organs. GPCRs modulate the opening and closing of the target ion channels either directly by binding them, or by releasing second messengers that activate these channels. As ions move across the membrane, the membrane potential is altered, which induces an appropriate response.
Sensory organs,...

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相关实验视频

Updated: Jul 12, 2026

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
11:51

Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Published on: April 27, 2018

活体中依赖金属离子的GFP排放通过将循环变异的绿色光蛋白与工程金属离子结合的卷轴-卷轴结合在一起.

Toshihisa Mizuno1, Kaori Murao, Yoichi Tanabe

  • 1Graduate School of Engineering, Nagoya Institute of Technology, Nagoya 466-8555, Japan. toshitcm@nitech.ac.jp

Journal of the American Chemical Society
|August 29, 2007
PubMed
概括
此摘要是机器生成的。

研究人员创造了一种融合蛋白质,该蛋白质将一种金属结合式卷轴连接到光蛋白质. 金属离子通过诱导卷轴结构触发光,从而使新的蛋白质设计成为可能.

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Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability
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Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability

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Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
10:41

Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity

Published on: February 17, 2017

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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence
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Engineering 'Golden' Fluorescence by Selective Pressure Incorporation of Non-canonical Amino Acids and Protein Analysis by Mass Spectrometry and Fluorescence

Published on: April 27, 2018

Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability
10:31

Residue-Specific Exchange of Proline by Proline Analogs in Fluorescent Proteins: How "Molecular Surgery" of the Backbone Affects Folding and Stability

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Using a GFP-tagged TMEM184A Construct for Confirmation of Heparin Receptor Identity
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Published on: February 17, 2017

科学领域:

  • 蛋白质工程是指蛋白质工程.
  • 生物化学 生物化学
  • 结构生物学 结构生物学

背景情况:

  • 金属离子协调对于蛋白质的结构和功能至关重要.
  • 设计具有可控制功能的新型蛋白质是生物技术的一个关键挑战.

研究的目的:

  • 设计一种融合蛋白质,其中金属离子与卷状卷状域结合,调节了一个循环 permuted 绿色光蛋白 (cpGFP) 的光.
  • 为了证明金属离子诱导的蛋白质折叠和功能在一个新设计的蛋白质系统.

主要方法:

  • 一个新设计的金属离子结合线圈 (IZ-HH) 的建造,使用胺残留物.
  • 一个循环 permuted GFP (cpGFP) 的工程和与IZ-HH域的融合.
  • 在大肠杆菌中融合蛋白的表达和对金属离子 (Cu2+, Zn2+) 的光反应的评估.

主要成果:

  • 在结合双价金属离子 (Cu2+,Ni2+,Zn2+) 时,IZ-HH卷状线圈形成了一个稳定的三元结构.
  • 聚变蛋白 (191) cpGFP(190) -IZ-HH在大肠杆菌细胞中表现出金属离子依赖的光.
  • 结果表明细胞膜成功通过,并通过细胞外金属离子诱导cpGFP光.

结论:

  • 开发了一种新的策略,用于设计金属离子响应蛋白,使用线圈-线圈开关.
  • 这种方法可以通过特定的金属离子调节蛋白质功能 (光).
  • 设计的系统显示了为在体外和体内生物应用创造多功能,非天然蛋白质的潜力.