Design and analysis of post-fusion 6-helix bundle of heptad repeat regions from Newcastle disease virus F protein. Protein Eng 2003 May;16(5):373-9
Date
06/27/2003Pubmed ID
12826729DOI
10.1093/protein/gzg041Scopus ID
2-s2.0-0038111976 (requires institutional sign-in at Scopus site) 19 CitationsAbstract
Fusion of paramyxovirus to the cell involves receptor binding of the HN glycoprotein and a number of conformational changes of F glycoprotein. The F protein is expressed as a homotrimer on the virus surface. In the present model, there are at least three conformations of F protein, i.e. native form, pre-hairpin intermediate and the post-fusion state. In the post-fusion state, the two highly conserved heptad repeat (HR) regions of F protein form a stable 6-helix coiled-coil bundle. However, no crystal structure is known for this state for the Newcastle disease virus, although the crystal structure of the F protein native form has been solved recently. Here we deployed an Escherichia coli in vitro expression system to engineer this 6-helix bundle by fusion of either the two HR regions (HR1, linker and HR2) or linking the 6-helix [3 x (HR1, linker and HR2)] together as a single chain. Subsequently, both of them form a stable 6-helix bundle in vitro judging by gel filtration and chemical cross-linking and the proteins show salient features of an alpha-helix structure. Crystals diffracting X-rays have been obtained from both protein preparations and the structure determination is under way. This method could be used for crystallization of the post-fusion state HR structures of other viruses.
Author List
Zhu J, Li P, Wu T, Gao F, Ding Y, Zhang CW, Rao Z, Gao GF, Tien PAuthor
Jieqing Zhu PhD Professor in the Biochemistry department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceChromatography, Gel
Circular Dichroism
Crystallization
Molecular Sequence Data
Newcastle disease virus
Protein Structure, Secondary
Viral Fusion Proteins
