title: Purification, crystallization and preliminary X-ray data collection of the N-terminal domain of the 26S proteasome regulatory subunit p27 and its complex with the ATPase domain of Rpt5 from Mus musculus.
abstract: The protein 26S proteasome regulatory subunit p27 is one of the four chaperones that help in the assembly of the 19S regulatory particle (RP) of the 26S proteasome. In the present work, the N-terminus of p27 (residues 1-128) from Mus musculus was cloned, expressed, purified and crystallized alone and in complex with the C-terminal ATPase domain of Rpt5 (residues 173-442). The crystals of p27((1-128)) diffracted to 1.7 Å resolution and belonged to space group P212121, with unit-cell parameters a = 26.79, b = 30.39, c = 145.06 Å. Resolution-dependent Matthews coefficient probability analysis suggested the presence of only one molecule per asymmetric unit, with 40.5% solvent content and a VM value of 2.02 Å(3) Da(-1). The crystal of the p27((1-128))-Rpt5((173-442)) complex diffracted to 4 Å resolution and belonged to space group P222, with unit-cell parameters a = 75.93, b = 76.08, c = 336.85 Å. The presence of four heterodimers in the asymmetric unit with 53.2% solvent content and a VM value of 2.63 Å(3) Da(-1) or five heterodimers in the asymmetric unit with 41.5% solvent content and a VM value of 2.10 Å(3) Da(-1) is assumed.
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, Nankai University, 94 Weijin Road, Tianjin 300071, People's Republic of China.
authors.forename: Wentao
authors.initials: W
authors.lastname: Diao
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, Nankai University, 94 Weijin Road, Tianjin 300071, People's Republic of China.
authors.forename: Xue
authors.initials: X
authors.lastname: Yang
authors.affiliation: State Key Laboratory of Medicinal Chemical Biology, Nankai University, 94 Weijin Road, Tianjin 300071, People's Republic of China.
authors.forename: Hao
authors.initials: H
authors.lastname: Zhou
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000302:isolation & purification*; D000818:Animals; D003460:Crystallization; D018360:Crystallography, X-Ray; D051379:Mice; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry* / Q000302:isolation & purification*; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry* / Q000302:isolation & purification*
chemical_list: D021122:Protein Subunits; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D000251:Adenosine Triphosphatases
keywords: 26S proteasome regulatory subunit p27; ATPase domain of Rpt5; Mus musculus; chaperones