PHOELIX: a package for semi-automated helical reconstruction.

We describe a set of procedures and algorithms which have been developed to provide an efficient and reliable method for reconstructing a three-dimensional density map from specimens with helical symmetry. These procedures build on the original MRC helical processing suite, with extensions principally developed using the SUPRIM image processing package. Actomyosin is used as a model specimen to demonstrate the utility of this repackaged and expanded set of routines. The time required to complete a three-dimensional map has been reduced from several weeks using traditional manual techniques to a few days. The increased signal/noise provided has allowed for the extraction of additional layer lines not previously identified by manual techniques.

[1]  R. Josephs,et al.  The reconstruction of helical particles with variable pitch. , 1988, Ultramicroscopy.

[2]  R A Milligan,et al.  Structural relationships of actin, myosin, and tropomyosin revealed by cryo-electron microscopy , 1987, The Journal of cell biology.

[3]  W. Kabsch,et al.  Atomic structure of the actin: DNase I complex , 1990, Nature.

[4]  D. DeRosier,et al.  The variable twist of actin and its modulation by actin-binding proteins , 1987, The Journal of cell biology.

[5]  J Jakana,et al.  Three-dimensional structure of a single filament in the Limulus acrosomal bundle: scruin binds to homologous helix-loop-beta motifs in actin , 1994, The Journal of cell biology.

[6]  D A Winkelmann,et al.  Three-dimensional structure of myosin subfragment-1: a molecular motor. , 1993, Science.

[7]  T. Baker,et al.  Structural studies of human alpha 2-macroglobulin: concordance between projected views obtained by negative-stain and cryoelectron microscopy. , 1991, Journal of structural biology.

[8]  Gebhard F. X. Schertler,et al.  Projection structure of rhodopsin , 1993, Nature.

[9]  E K Fram,et al.  Reconstruction of imperfectly ordered zinc-induced tubulin sheets using cross-correlation and real space averaging. , 1981, Ultramicroscopy.

[10]  M. Whittaker,et al.  Molecular structure of F-actin and location of surface binding sites , 1990, Nature.

[11]  D. DeRosier,et al.  The Fourier transform of actin and other helical systems with cumulative random angular disorder , 1982 .

[12]  K. Downing,et al.  Assessment of resolution in biological electron crystallography. , 1992, Ultramicroscopy.

[13]  Marin van Heel,et al.  Invariant recognition of molecular projections in vitreous ice preparations , 1992 .

[14]  R. Craig,et al.  Three-dimensional reconstruction of thin filaments decorated with a Ca2+-regulated myosin. , 1982, Journal of molecular biology.

[15]  E. Egelman,et al.  An algorithm for straightening images of curved filamentous structures. , 1986, Ultramicroscopy.

[16]  R Henderson,et al.  Image contrast in high-resolution electron microscopy of biological macromolecules: TMV in ice. , 1992, Ultramicroscopy.

[17]  U. Aebi,et al.  The Three‐Dimensional Structure of the Actin Filament Revisited a , 1986, Annals of the New York Academy of Sciences.

[18]  R A Milligan,et al.  Structure of the actin-myosin complex and its implications for muscle contraction. , 1993, Science.

[19]  D. DeRosier,et al.  Three-dimensional reconstruction of F-actin, thin filaments and decorated thin filaments. , 1970, Journal of molecular biology.

[20]  W. Kabsch,et al.  Atomic model of the actin filament , 1990, Nature.

[21]  J. Hanson.  Axial Period of Actin Filaments , 1967, Nature.

[22]  D G Morgan,et al.  Processing images of helical structures: a new twist. , 1992, Ultramicroscopy.

[23]  D. DeRosier,et al.  Determination of the alpha-actinin-binding site on actin filaments by cryoelectron microscopy and image analysis , 1994, The Journal of cell biology.

[24]  A. Klug,et al.  Three-dimensional image reconstructions of the contractile tail of T4 bacteriophage. , 1975, Journal of molecular biology.

[25]  N. Unwin.  Nicotinic acetylcholine receptor at 9 A resolution. , 1993, Journal of molecular biology.

[26]  C. Toyoshima,et al.  Three-dimensional image analysis of the complex of thin filaments and myosin molecules from skeletal muscle. IV. Reconstitution from minimal- and high-dose images of the actin-tropomyosin-myosin subfragment-1 complex. , 1985, Journal of biochemistry.

[27]  L. Amos,et al.  A new model for the geometry of the binding of myosin crossbridges to muscle thin filaments. , 1981, Journal of molecular biology.

[28]  D. DeRosier,et al.  Reconstruction of three-dimensional images from electron micrographs of structures with helical symmetry. , 1970, Journal of molecular biology.

[29]  A. Klug,et al.  Three-dimensional image reconstruction of actin-tropomyosin complex and actin-tropomyosin-troponin T-troponin I complex. , 1975, Journal of molecular biology.