Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages

Investigations into protein quaternary structure can lead to deeper insight into the fundamentals governing both protein folding and protein-protein interactions. In addition they can provide a foundation for the eventual rational design of novel complex protein architectures. A maxi-ferritin, bacte...

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Main Author: Zhang, Yu
Other Authors: Brendan Patrick Orner
Format: Theses and Dissertations
Language:English
Published: 2012
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Online Access:https://hdl.handle.net/10356/48015
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-480152023-02-28T23:43:13Z Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages Zhang, Yu Brendan Patrick Orner School of Physical and Mathematical Sciences DRNTU::Science::Chemistry::Biochemistry Investigations into protein quaternary structure can lead to deeper insight into the fundamentals governing both protein folding and protein-protein interactions. In addition they can provide a foundation for the eventual rational design of novel complex protein architectures. A maxi-ferritin, bacterioferritin from E. coli (BFR), and a mini-ferritin, DNA-binding protein from starved cells (DPS), despite their similar four-helix bundle tertiary structure, assemble into quaternary structure with different symmetries, octahedral and tetrahedral and oligomerization states, 24-mer and 12-mer, respectively. To understand how these two structurally analogous proteins assemble into nano-structures with different sizes and shapes, both proteins were chosen as the basis for a mutagenesis study to investigate the importance of key amino acid residues, located at symmetry-related protein-protein interfaces, in controlling protein stability and self-assembly. Several mutants were designed for each protein through simple inspection and computational analysis, synthesized and subjected to different chemical and biophysical methods to determine their thermal stability, self-assemble ability and structure. The data indicate that many of these residues may be hot spot residues. Several mutants were observed to completely shut down detectable solution formation of 24-mer, favoring a cooperatively folded dimer, suggesting that they may be oligomerization “switch residues”. This investigation into the structure and energetics of these self-assembling nano-cage proteins not only can act as a jumping off point for the eventual design of novel protein nano-structures and their applications, but it can also help to understand the role that structure plays in the function of these important classes of proteins. DOCTOR OF PHILOSOPHY (SPMS) 2012-02-14T00:56:48Z 2012-02-14T00:56:48Z 2011 2011 Thesis Zhang, Y. (2011). Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages. Doctoral thesis, Nanyang Technological University, Singapore. https://hdl.handle.net/10356/48015 10.32657/10356/48015 en 191 p. application/pdf
institution Nanyang Technological University
building NTU Library
continent Asia
country Singapore
Singapore
content_provider NTU Library
collection DR-NTU
language English
topic DRNTU::Science::Chemistry::Biochemistry
spellingShingle DRNTU::Science::Chemistry::Biochemistry
Zhang, Yu
Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
description Investigations into protein quaternary structure can lead to deeper insight into the fundamentals governing both protein folding and protein-protein interactions. In addition they can provide a foundation for the eventual rational design of novel complex protein architectures. A maxi-ferritin, bacterioferritin from E. coli (BFR), and a mini-ferritin, DNA-binding protein from starved cells (DPS), despite their similar four-helix bundle tertiary structure, assemble into quaternary structure with different symmetries, octahedral and tetrahedral and oligomerization states, 24-mer and 12-mer, respectively. To understand how these two structurally analogous proteins assemble into nano-structures with different sizes and shapes, both proteins were chosen as the basis for a mutagenesis study to investigate the importance of key amino acid residues, located at symmetry-related protein-protein interfaces, in controlling protein stability and self-assembly. Several mutants were designed for each protein through simple inspection and computational analysis, synthesized and subjected to different chemical and biophysical methods to determine their thermal stability, self-assemble ability and structure. The data indicate that many of these residues may be hot spot residues. Several mutants were observed to completely shut down detectable solution formation of 24-mer, favoring a cooperatively folded dimer, suggesting that they may be oligomerization “switch residues”. This investigation into the structure and energetics of these self-assembling nano-cage proteins not only can act as a jumping off point for the eventual design of novel protein nano-structures and their applications, but it can also help to understand the role that structure plays in the function of these important classes of proteins.
author2 Brendan Patrick Orner
author_facet Brendan Patrick Orner
Zhang, Yu
format Theses and Dissertations
author Zhang, Yu
author_sort Zhang, Yu
title Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
title_short Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
title_full Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
title_fullStr Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
title_full_unstemmed Mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
title_sort mutagenesis studies to determine the key interfacial amino acid residues that govern the self-assembly of the ferritin protein cages
publishDate 2012
url https://hdl.handle.net/10356/48015
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