Crystallization of Saccharomyces cerevisiae aminopeptidase 1, the major cargo protein of the Cvt pathway.
The vacuole hydrolase aminopeptidase 1 (Ape1) is a cargo protein transported to the vacuole by the cytosol-to-vacuole targeting (Cvt) pathway during conditions of growth and by autophagy during conditions of starvation. After transport to the vacuole, Ape1 is processed into mature Ape1 (mApe1). mApe1 has been expressed, purified and crystallized in two crystal forms. Form I belongs to space group P2(1), with unit-cell parameters a = 120.6, b = 219.5, c = 133.1 A, beta = 116.5 degrees. Form II belongs to space group R3, with unit-cell parameters a = 141.2, c = 349.4 A. Diffraction data were collected from these crystals to a resolution of 2.5 A for form I and 1.83 A for form II. Self-rotation functions and the volume-to-weight ratio values suggest that forms I and II contain 12 and four mApe1 molecules per asymmetric unit, respectively, and that mApe1 exists as a tetrahedral dodecamer in both crystal forms.
Adachi
Wakana
W
Department of Structural Biology, Graduate School of Pharmaceutical Sciences, Hokkaido University, N-12, W-6, Kita-ku, Sapporo 060-0812, Japan.
Suzuki
Nobuo N
NN
Fujioka
Yuko
Y
Suzuki
Kuninori
K
Ohsumi
Yoshinori
Y
Inagaki
Fuyuhiko
F
D000626:Aminopeptidases / Q000737:chemistry* / Q000502:physiology; D003460:Crystallization; D003600:Cytosol / Q000201:enzymology* / Q000502:physiology; D007527:Isoenzymes / Q000737:chemistry / Q000502:physiology; D008970:Molecular Weight; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000502:physiology; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000502:physiology; D015398:Signal Transduction / Q000502:physiology*; D014617:Vacuoles / Q000201:enzymology* / Q000502:physiology; D033921:Vesicular Transport Proteins / Q000737:chemistry* / Q000502:physiology
D007527:Isoenzymes; D029701:Saccharomyces cerevisiae Proteins; D033921:Vesicular Transport Proteins; D000626:Aminopeptidases; C019689:APE1 protein, S cerevisiae