Purification, crystallization and preliminary X-ray diffraction analysis of the non-ATPase subunit Nas6 in complex with the ATPase subunit Rpt3 of the 26S proteasome from Saccharomyces cerevisiae.
The non-ATPase subunit Nas6, which is the human orthologue of gankyrin, was co-expressed with the C-terminal domain of the ATPase subunit Rpt3 of the yeast 26S proteasome in Escherichia coli, purified to near-homogeneity and crystallized using the hanging-drop vapour-diffusion method. The protein crystallized in space group P2(1), with unit-cell parameters a = 60.38, b = 100.22, c = 72.20 A, beta = 94.70 degrees and with three Nas6-Rpt3C molecules per asymmetric unit. The crystal diffracted to beyond 2.2 A resolution using synchrotron radiation.
Nakamura
Yoshihiro
Y
RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi, Yokohama 230-0045, Japan.
Umehara
Takashi
T
Tanaka
Akiko
A
Horikoshi
Masami
M
Padmanabhan
Balasundaram
B
Yokoyama
Shigeyuki
S
D000251:Adenosine Triphosphatases / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000302:isolation & purification; D006801:Humans; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry* / Q000302:isolation & purification; D021122:Protein Subunits / Q000737:chemistry* / Q000302:isolation & purification; D011518:Proto-Oncogene Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000302:isolation & purification; D014961:X-Ray Diffraction
D004268:DNA-Binding Proteins; C114551:NAS6 protein, S cerevisiae; C403591:PSMD10 protein, human; D021122:Protein Subunits; D011518:Proto-Oncogene Proteins; D029701:Saccharomyces cerevisiae Proteins; D046988:Proteasome Endopeptidase Complex; C061553:ATP dependent 26S protease; D000251:Adenosine Triphosphatases; C084493:RPT1 protein, S cerevisiae