title: Expression, purification, crystallization and preliminary crystallographic analysis of hepatitis B virus core protein dimerized via a peptide linker containing an EGFP insertion.
abstract: 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 F432, 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.
authors.affiliation: Faculty of Pharmaceutical Sciences, Chiba Institute of Science, 15-8 Shiomi-cho, Chosi, Chiba 288-0025, Japan.
authors.forename: Masaki
authors.initials: M
authors.lastname: Kikuchi
authors.forename: Shinichiro
authors.initials: S
authors.lastname: Iwabuchi
authors.forename: Tatsuhiko
authors.initials: T
authors.lastname: Kikkou
authors.forename: Keiichi
authors.initials: K
authors.lastname: Noguchi
authors.forename: Masafumi
authors.initials: M
authors.lastname: Odaka
authors.forename: Masafumi
authors.initials: M
authors.lastname: Yohda
authors.forename: Masaaki
authors.initials: M
authors.lastname: Kawata
authors.forename: Chikara
authors.initials: C
authors.lastname: Sato
authors.forename: Osamu
authors.initials: O
authors.lastname: Matsumoto
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D015967:Gene Expression Regulation, Viral; D049452:Green Fluorescent Proteins / Q000235:genetics*; D006512:Hepatitis B Core Antigens / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006515:Hepatitis B virus / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D016254:Mutagenesis, Insertional / Q000235:genetics / Q000276:immunology; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics*; D055503:Protein Multimerization / Q000502:physiology; D014758:Viral Core Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification
chemical_list: D006512:Hepatitis B Core Antigens; D010446:Peptide Fragments; D014758:Viral Core Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: enhanced green fluorescent protein; fusion protein; hepatitis B virus
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