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Vac8p is a vacuolar protein that plays pivotal roles in both vacuole inheritance and the formation of nucleus vacuolejunction (NVJ) in yeast. The Vac8p directly interacts with Atg13p, a component of the autophagy machinery, and mediatescytoplasm-to-vacuole targeting (Cvt) pathway, resulting in the maturation of aminopeptidase I (Ape1p). Here, we coexpressedand purified Saccharomyces cerevisiae Vac8p complexed with Atg13p in Escherichia coli bacteria cells, andcrystallized the complex proteins under the condition of 25% (v/v) PEG 400, 100 mM Tris pH 8.5, 2% (v/v) Ethylene glycol,2% (w/v) PEG 3350, 1.5% (w/v) PEG 20000, 5 mM DTT at 293K. X-ray diffraction data of the crystals were collected to2.9 A resolution at the synchrotron radiation. The crystals belong to the orthorhombic space group P212121 with unit cellparameters a = 62.7 A, b = 92.4 A, and c = 139.9 A. The asymmetric unit contains one Vac8p-Atg13p heterodimer with acorresponding VM of 2.92 A3 Da-1 and solvent content of 57.8%.
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참고문헌 (16)
참고문헌 신청
Funakoshi, T / 1997 / Analyses of APG13 gene involved in autophagy in yeast, Saccharomyces cerevisiae / Gene 192:207~213
Jeong, H / 2017 / Mechanistic insight into the nucleus-vacuole junction based on the Vac8p-Nvj1p crystal structure / Proc Natl Acad Sci U S A 114:E4539~E4548
Klionsky, D. J / 1992 / Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway / J Cell Biol 119:287~299
Klionsky, D. J / 1990 / The fungal vacuole : composition, function, and biogenesis / Microbiol Rev 54:266~292
Ishikawa, K / 2003 / Identification of an organelle-specific myosin V receptor / J Cell Biol 160:887~897
Jeong, H / 2017 / Mechanistic insight into the nucleus-vacuole junction based on the Vac8p-Nvj1p crystal structure / Proc Natl Acad Sci U S A 114:E4539~E4548
Klionsky, D. J / 1992 / Aminopeptidase I of Saccharomyces cerevisiae is localized to the vacuole independent of the secretory pathway / J Cell Biol 119:287~299
Klionsky, D. J / 1990 / The fungal vacuole : composition, function, and biogenesis / Microbiol Rev 54:266~292
Ishikawa, K / 2003 / Identification of an organelle-specific myosin V receptor / J Cell Biol 160:887~897
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