Helical structure of unidirectionally shadowed metal replicas of cyanide hydratase from Gloeocercospora sorghi.
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Helical structure of unidirectionally shadowed metal replicas of cyanide hydratase from Gloeocercospora sorghi. / Woodward, J D; Weber, Beatrice; Scheffer, M P; Benedik, M J; Hoenger, A; Sewell, B T.
In: J STRUCT BIOL, Vol. 161, No. 2, 2, 2008, p. 111-119.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Helical structure of unidirectionally shadowed metal replicas of cyanide hydratase from Gloeocercospora sorghi.
AU - Woodward, J D
AU - Weber, Beatrice
AU - Scheffer, M P
AU - Benedik, M J
AU - Hoenger, A
AU - Sewell, B T
PY - 2008
Y1 - 2008
N2 - The helical filaments of the cyanide hydratase from Gloeocercospora sorghi have been reconstructed in three dimensions from freeze dried, unidirectionally shadowed specimens using iterative real-space helical reconstruction. The average power spectrum of all selected images has three clear reflections on different layer lines. The reconstruction is complicated by the fact that three possible indexing schemes are possible and reconstructions using the starting symmetries based on each of these indexing schemes converge on three-dimensional volumes which appear plausible. Because only one side is visible in shadowed specimens, it is necessary to examine the phases from a single filament by cryo-electron microscopy in order to make an unequivocal assignment of the symmetry. Because of the novel nature of the reconstruction method used here, conventional cryo-EM methods were also used to determine a second reconstruction, allowing us to make comparisons between the two. The filament is shown to have a left-handed one-start helix with D(1) symmetry, 5.46 dimers per turn and a pitch of 7.15nm. The reconstruction suggests the presence of an interaction across the groove not previously seen in nitrilase helical fibres.
AB - The helical filaments of the cyanide hydratase from Gloeocercospora sorghi have been reconstructed in three dimensions from freeze dried, unidirectionally shadowed specimens using iterative real-space helical reconstruction. The average power spectrum of all selected images has three clear reflections on different layer lines. The reconstruction is complicated by the fact that three possible indexing schemes are possible and reconstructions using the starting symmetries based on each of these indexing schemes converge on three-dimensional volumes which appear plausible. Because only one side is visible in shadowed specimens, it is necessary to examine the phases from a single filament by cryo-electron microscopy in order to make an unequivocal assignment of the symmetry. Because of the novel nature of the reconstruction method used here, conventional cryo-EM methods were also used to determine a second reconstruction, allowing us to make comparisons between the two. The filament is shown to have a left-handed one-start helix with D(1) symmetry, 5.46 dimers per turn and a pitch of 7.15nm. The reconstruction suggests the presence of an interaction across the groove not previously seen in nitrilase helical fibres.
M3 - SCORING: Zeitschriftenaufsatz
VL - 161
SP - 111
EP - 119
JO - J STRUCT BIOL
JF - J STRUCT BIOL
SN - 1047-8477
IS - 2
M1 - 2
ER -
