Crystallization of Saccharomyces cerevisiae alpha-mannosidase, a cargo protein of the Cvt pathway.
Saccharomyces cerevisiae alpha-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 P2(1), with unit-cell parameters a = 145.7, b = 127.7, c = 164.0 A, beta = 101.5 degrees . Form II belongs to space group I222 or I2(1)2(1)2(1), with unit-cell parameters a = 127.9, b = 163.7, c = 291.5 A. Diffraction data were collected from these crystals to a resolution of 3.3 A for form I and of 2.6 A for form II using synchrotron radiation.
Watanabe
Yasunori
Y
Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, N-12, W-6, Kita-ku, Sapporo 060-0812, Japan.
Noda
Nobuo N
NN
Honbou
Kazuya
K
Suzuki
Kuninori
K
Sakai
Yasuyoshi
Y
Ohsumi
Yoshinori
Y
Inagaki
Fuyuhiko
F
D001343:Autophagy / Q000502:physiology; D003460:Crystallization; D018360:Crystallography, X-Ray; D003625:Data Collection; D010843:Pichia / Q000235:genetics; D021381:Protein Transport; D012399:Rotation; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism*; D013223:Statistics as Topic; D017356:Synchrotrons; D014617:Vacuoles / Q000378:metabolism; D033921:Vesicular Transport Proteins / Q000378:metabolism*; D014961:X-Ray Diffraction; D043323:alpha-Mannosidase / Q000302:isolation & purification / Q000378:metabolism*
D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; D043323:alpha-Mannosidase