Open AccessCrystallization of Saccharomyces cerevisiae α‐mannosidase, a cargo protein of the Cvt pathway
Author(s) -
Watanabe Yasunori,
Noda Nobuo N.,
Honbou Kazuya,
Suzuki Kuninori,
Sakai Yasuyoshi,
Ohsumi Yoshinori,
Inagaki Fuyuhiko
Publication year - 2009
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309109015826
Subject(s) - mannosidase , saccharomyces cerevisiae , crystallization , chemistry , protein crystallization , biochemistry , chemical engineering , yeast , enzyme , organic chemistry , engineering
Saccharomyces cerevisiae α‐mannosidase (Ams1) is a cargo protein that is transported to the vacuole by the cytoplasm‐to‐vacuole targeting (Cvt) pathway during conditions of growth and by autophagy during conditions of starvation. After transport to the vacuole, Ams1 functions as a resident hydrolase. Ams1 has been overexpressed in the methylotrophic yeast Pichia pastoris , purified and crystallized in two crystal forms. Form I belongs to space group P 2 1 , with unit‐cell parameters a = 145.7, b = 127.7, c = 164.0 Å, β = 101.5°. Form II belongs to space group I 222 or I 2 1 2 1 2 1 , with unit‐cell parameters a = 127.9, b = 163.7, c = 291.5 Å. Diffraction data were collected from these crystals to a resolution of 3.3 Å for form I and of 2.6 Å for form II using synchrotron radiation.