Screening for interaction domain in vault nanocapsules

Protein nanocapsule is gaining popularity as therapeutic delivery vehicle. pH-controlled molecular release from the protein nanocapsule is particularly interesting. Vault is a naturally assembled hollow protein nanocapsule (72 x 41 x 41 nm3) which is composed of major vault protein (MVP) domain that...

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Main Author: Vania, Maya
Other Authors: Lim Sierin
Format: Final Year Project
Language:English
Published: 2010
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Online Access:http://hdl.handle.net/10356/39606
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Institution: Nanyang Technological University
Language: English
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spelling sg-ntu-dr.10356-396062023-03-03T15:40:22Z Screening for interaction domain in vault nanocapsules Vania, Maya Lim Sierin School of Chemical and Biomedical Engineering DRNTU::Engineering::Bioengineering Protein nanocapsule is gaining popularity as therapeutic delivery vehicle. pH-controlled molecular release from the protein nanocapsule is particularly interesting. Vault is a naturally assembled hollow protein nanocapsule (72 x 41 x 41 nm3) which is composed of major vault protein (MVP) domain that interacts with C-terminus of vault poly(ADP-ribose)-polymerase (VPARP), namely MVP-minimal interaction domain (mINT). The interaction between MVP and mINT is important in engineering the non-native pH-dependent interaction functionality onto the protein nanocapsule. We attempted to identify specific domains that are involved in the interactions between MVP and mINT. From the electron difference map, mINT is hypothesized to associate at the N-terminal MVP at domain 2-5. We constructed four plasmids in E. coli expression vector pET-11a and pET-28a encoding MVP domain 3-4, 3-5, 2-5 and mCherry fused with mINT. These genes were overexpressed to produce protein fractions for validation of interactions. Analysis by SDS-PAGE revealed that majority of protein mCherry-mINT was found in soluble fraction while MVP domain 2-5 was largely presented in pellet form. The use of polyhistidine tag improved the expression of MVP domain 3-5. However, MVP domain 3-4 was not expressed in sufficient quantity to be detected. Bachelor of Engineering (Chemical and Biomolecular Engineering) 2010-06-01T06:50:54Z 2010-06-01T06:50:54Z 2010 2010 Final Year Project (FYP) http://hdl.handle.net/10356/39606 en Nanyang Technological University 76 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::Engineering::Bioengineering
spellingShingle DRNTU::Engineering::Bioengineering
Vania, Maya
Screening for interaction domain in vault nanocapsules
description Protein nanocapsule is gaining popularity as therapeutic delivery vehicle. pH-controlled molecular release from the protein nanocapsule is particularly interesting. Vault is a naturally assembled hollow protein nanocapsule (72 x 41 x 41 nm3) which is composed of major vault protein (MVP) domain that interacts with C-terminus of vault poly(ADP-ribose)-polymerase (VPARP), namely MVP-minimal interaction domain (mINT). The interaction between MVP and mINT is important in engineering the non-native pH-dependent interaction functionality onto the protein nanocapsule. We attempted to identify specific domains that are involved in the interactions between MVP and mINT. From the electron difference map, mINT is hypothesized to associate at the N-terminal MVP at domain 2-5. We constructed four plasmids in E. coli expression vector pET-11a and pET-28a encoding MVP domain 3-4, 3-5, 2-5 and mCherry fused with mINT. These genes were overexpressed to produce protein fractions for validation of interactions. Analysis by SDS-PAGE revealed that majority of protein mCherry-mINT was found in soluble fraction while MVP domain 2-5 was largely presented in pellet form. The use of polyhistidine tag improved the expression of MVP domain 3-5. However, MVP domain 3-4 was not expressed in sufficient quantity to be detected.
author2 Lim Sierin
author_facet Lim Sierin
Vania, Maya
format Final Year Project
author Vania, Maya
author_sort Vania, Maya
title Screening for interaction domain in vault nanocapsules
title_short Screening for interaction domain in vault nanocapsules
title_full Screening for interaction domain in vault nanocapsules
title_fullStr Screening for interaction domain in vault nanocapsules
title_full_unstemmed Screening for interaction domain in vault nanocapsules
title_sort screening for interaction domain in vault nanocapsules
publishDate 2010
url http://hdl.handle.net/10356/39606
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