fulltext.study @t Gmail

Oligomerization state of the multimerization domain of Nipah virus phosphoprotein

Paper ID Volume ID Publish Year Pages File Format Full-Text
34542 45033 2013 5 PDF Available
Title
Oligomerization state of the multimerization domain of Nipah virus phosphoprotein
Abstract

•Nipah virus (NiV) phosphoprotein multimerization domain (PMD) was expressed in E. coli.•The PMD protein was purified using IMAC which yielded a purity of 97%.•Chemical cross-linking and mass spectrometry revealed that the PMD formed tetramer.•Circular dichroism analysis revealed that NiV PMD contains high percentage of α-helical structure.

Oligomerization of the phospho (P) proteins of paramyxoviruses plays important roles in the viral genome replication and transcription. The multimerization domain of the P proteins (PMD) was found to be responsible and sufficient for the oligomerization process. However, the oligomeric status of Nipah virus (NiV) P protein is still unknown. Therefore, the aim of this study was to produce the PMD region of NiV in bacteria in order to determine the oligomeric status of the P protein. The NiV PMD region was cloned into pTrcHis2-TOPO vector and expressed in Escherichia coli. SDS-PAGE and Western blot analysis confirmed the protein expression upon IPTG induction at 37 °C for 5 h. Immobilized metal affinity chromatography (IMAC) purification was developed to purify the recombinant histidine-tagged NiV PMD which yielded a purity of 97%. Chemical cross-linking revealed that the PMD of NiV formed tetramer. The stability of the oligomer was studied with dynamic light scattering while mass spectrometry confirmed the exact molecular masses of the cross-linked products. Circular dichroism analysis revealed that the NiV PMD contained high α-helical content (55%) suggesting the presence of a coiled coil structure.

Keywords
Nipah virus; Phosphoprotein; Multimerization domain; Oligomerization; Coiled coil structure
First Page Preview
Oligomerization state of the multimerization domain of Nipah virus phosphoprotein
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us
Publisher
Database: Elsevier - ScienceDirect
Journal: Process Biochemistry - Volume 48, Issue 10, October 2013, Pages 1476–1480
Authors
, , , , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us