The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer
碩士 === 國立交通大學 === 生物科技學系 === 99 === ―How does a protein fold?‖ This has been questioned for long period. Studying on a single-domain protein can be a remark to understand the protein folding mechanism in general. Recently, we folded the Protein G B1 Domain (PGB1) by an over-critical folding process,...
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ndltd-TW-099NCTU51111332015-10-13T20:37:27Z http://ndltd.ncl.edu.tw/handle/60105143968599265338 The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer 以螢光共振能量轉移揭示蛋白質PGB1分子內摺疊機制 鍾偉賢 碩士 國立交通大學 生物科技學系 99 ―How does a protein fold?‖ This has been questioned for long period. Studying on a single-domain protein can be a remark to understand the protein folding mechanism in general. Recently, we folded the Protein G B1 Domain (PGB1) by an over-critical folding process, which was developed by our lab, and studied the conformational changes in each folding state. PGB1 is a small protein with 56 residues, and is an IgG-binding domain of protein G. Because PGB1 contains the basic folding elements in a short sequence of amino acid without disulfide bonds, making it an excellent model for protein folding studies. The intrinsic Trp43 fluorescence of PGB1 and the acrylamide-quenching fluorescence showed that the PGB1 folding is a two-state reaction. However, the molecular diameter changes during the folding process indicated a folding intermediate exists in the PGB1 folding process. The FRET experiment, with Trp/IAEDANS as fluorophore pairs, showed that the β-hairpin 2 attaches to the α-helix long before the β-hairpin 1 formation. All the results together suggest the PGB1 folding process is a multi-state reaction. Overall, we were successful to reveal the folding process of PGB1 by FRET analysis. 張家靖 2011 學位論文 ; thesis 77 zh-TW |
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碩士 === 國立交通大學 === 生物科技學系 === 99 === ―How does a protein fold?‖ This has been questioned for long period. Studying on a single-domain protein can be a remark to understand the protein folding mechanism in general. Recently, we folded the Protein G B1 Domain (PGB1) by an over-critical folding process, which was developed by our lab, and studied the conformational changes in each folding state. PGB1 is a small protein with 56 residues, and is an IgG-binding domain of protein G. Because PGB1 contains the basic folding elements in a short sequence of amino acid without disulfide bonds, making it an excellent model for protein folding studies. The intrinsic Trp43 fluorescence of PGB1 and the acrylamide-quenching fluorescence showed that the PGB1 folding is a two-state reaction. However, the molecular diameter changes during the folding process indicated a folding intermediate exists in the PGB1 folding process. The FRET experiment, with Trp/IAEDANS as fluorophore pairs, showed that the β-hairpin 2 attaches to the α-helix long before the β-hairpin 1 formation. All the results together suggest the PGB1 folding process is a multi-state reaction. Overall, we were successful to reveal the folding process of PGB1 by FRET analysis.
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張家靖 |
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張家靖 鍾偉賢 |
author |
鍾偉賢 |
spellingShingle |
鍾偉賢 The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer |
author_sort |
鍾偉賢 |
title |
The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer |
title_short |
The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer |
title_full |
The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer |
title_fullStr |
The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer |
title_full_unstemmed |
The Folding Mechanism of Protein G B1 Domain Revealed by Fluorescence Resonance Energy Transfer |
title_sort |
folding mechanism of protein g b1 domain revealed by fluorescence resonance energy transfer |
publishDate |
2011 |
url |
http://ndltd.ncl.edu.tw/handle/60105143968599265338 |
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