Open AccessExpression, purification, crystallization and preliminary crystallographic analysis of hepatitis B virus core protein dimerized via a peptide linker containing an EGFP insertion
Author(s) -
Kikuchi Masaki,
Iwabuchi Shinichiro,
Kikkou Tatsuhiko,
Noguchi Keiichi,
Odaka Masafumi,
Yohda Masafumi,
Kawata Masaaki,
Sato Chikara,
Matsumoto Osamu
Publication year - 2013
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309113019957
Subject(s) - linker , crystallization , crystallography , peptide , icosahedral symmetry , heterologous , escherichia coli , chemistry , octahedron , crystal structure , biochemistry , gene , organic chemistry , computer science , operating system
Virus‐like particles (VLPs) have many potentially useful applications. The core proteins of human hepatitis B virus self‐assemble into icosahedral VLPs. As previously reported, core protein dimers (CPDs), produced by connecting two core proteins via a peptide linker, can also assemble into VLPs. CPDs in which heterologous proteins were connected to the C‐terminus (CPD1) were found to rearrange into symmetrical octahedra during crystallization. In this study, a heterologous protein was inserted into the peptide linker of the CPD (CPD2). CPD2 was expressed in Escherichia coli , assembled into VLPs, purified and crystallized. A single crystal diffracted to 2.8 Å resolution and belonged to the cubic space group F 432, with unit‐cell parameters a = b = c = 218.6 Å. Single‐crystal analysis showed that CPD1 and CPD2 rearranged into the same octahedral organization in a crystallization solution.