The rat peptidylarginine deiminase-encoding gene: structural analysis and the 5′-flanking sequence

Peptidylarginine deiminase (PAD) is a Ca2+-dependent enzyme that catalyzes the conversion of protein arginine residues to citrulline. Protein citrullination by PAD confers large structural and mechanical effects on the target proteins by altering intermolecular and intramolecular ionic or hydrophobic interactions. Five paralogous genes (PADI1 - 4 and 6) on human chromosome 1p35-36 encode the human PAD isozymes. Among the PADs, PAD3 shows the highest substrate specificity for synthetic and natural substrate. S100A3 is an EF-hand-type Ca2+-binding S100 protein family member that colocalizes with PAD3 in hair cuticular cells. PAD3 converts a symmetric pair of Arg51 residues on an S100A3 dimer surface to citrullines, causing assembly of a homotetramer, but does not convert other arginines. Although this specific citrullination is largely affected by the formation of two intramolecular disulfides in S100A3, it is not clear how the sheltered Arg51 residues is recognized by PAD3. We are aiming structural analysis of the substrate bound forms to elucidate structural factor that PAD3 recognize Arg51 residues only. Although X-ray crystal structures of the PAD4 isozyme and its complexes with substrate peptides have been reported, structural analysis of other PAD isozymes has not yet been conducted. To obtain the crystals of the substrate-complex, we prepared the C646A mutant and the other inactive mutants (D350A, H470A, D472A) of PAD3. We determined the crystal structures of wild-type PAD3, at first. Then, we have tried to determine the structure of the substrate complex with the mutants of PAD3. However, the solved structures did not contain the substrate at present stage. From our structural analysis, only the crystals of C646A were belonged to different crystal system from the others, and its difference didn't relate to their crystallization condition. In this conference, we discuss the origin(s) of the differences in the crystal system of C646 from the others.

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The gene encoding mouse lymphocyte antigen Ly-49: structural analysis and the 5'-flanking sequence

Gene ◽

10.1016/0378-1119(93)90489-p ◽

1993 ◽

Vol 136 (1-2) ◽

pp. 329-331 ◽

Cited By ~ 25

Author(s):

Kubo Sachiho ◽

Itoh Yohjiro ◽

Ishikawa Naoshi ◽

Nagasawa Ryuji ◽

Mitarai Tetsuya ◽

...

Keyword(s):

Structural Analysis ◽

Flanking Sequence ◽

Gene Encoding ◽

Lymphocyte Antigen ◽

Mouse Lymphocyte

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Structural analysis of the surface-layer protein of spirillum serpens by high-resolution electron microscopy

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100077268 ◽

1983 ◽

Vol 41 ◽

pp. 738-739

Author(s):

W. H. Wu ◽

R. M. Glaeser

Keyword(s):

Electron Microscopy ◽

Surface Layer ◽

Structural Analysis ◽

High Resolution ◽

Low Dose ◽

High Resolution Electron Microscopy ◽

Optical Diffraction ◽

Surface Layer Protein ◽

Morphological Unit ◽

Resolution Electron

Spirillum serpens possesses a surface layer protein which exhibits a regular hexagonal packing of the morphological subunits. A morphological model of the structure of the protein has been proposed at a resolution of about 25 Å, in which the morphological unit might be described as having the appearance of a flared-out, hollow cylinder with six ÅspokesÅ at the flared end. In order to understand the detailed association of the macromolecules, it is necessary to do a high resolution structural analysis. Large, single layered arrays of the surface layer protein have been obtained for this purpose by means of extensive heating in high CaCl2, a procedure derived from that of Buckmire and Murray. Low dose, low temperature electron microscopy has been applied to the large arrays.As a first step, the samples were negatively stained with neutralized phosphotungstic acid, and the specimens were imaged at 40,000 magnification by use of a high resolution cold stage on a JE0L 100B. Low dose images were recorded with exposures of 7-9 electrons/Å2. The micrographs obtained (Fig. 1) were examined by use of optical diffraction (Fig. 2) to tell what areas were especially well ordered.

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RNA polymerase: Preparation and structural analysis of helical polymers

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010011091x ◽

1984 ◽

Vol 42 ◽

pp. 152-153

Author(s):

E. Loren Buhle ◽

Pamela Rew ◽

Ueli Aebi

Keyword(s):

Structural Analysis ◽

Rna Polymerase ◽

Molecular Level ◽

X Ray Diffraction ◽

Helical Polymers ◽

X Ray ◽

E Coli ◽

The Past ◽

Ordered Arrays ◽

Low Ionic Strength

While DNA-dependent RNA polymerase represents one of the key enzymes involved in transcription and ultimately in gene expression in procaryotic and eucaryotic cells, little progress has been made towards elucidation of its 3-D structure at the molecular level over the past few years. This is mainly because to date no 3-D crystals suitable for X-ray diffraction analysis have been obtained with this rather large (MW ~500 kd) multi-subunit (α2ββ'ζ). As an alternative, we have been trying to form ordered arrays of RNA polymerase from E. coli suitable for structural analysis in the electron microscope combined with image processing. Here we report about helical polymers induced from holoenzyme (α2ββ'ζ) at low ionic strength with 5-7 mM MnCl2 (see Fig. 1a). The presence of the ζ-subunit (MW 86 kd) is required to form these polymers, since the core enzyme (α2ββ') does fail to assemble into such structures under these conditions.

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Interpreting HVEM of muscle-cell impulse networks

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s042482010012103x ◽

1992 ◽

Vol 50 (1) ◽

pp. 126-127

Author(s):

Paul DeCosta ◽

Kyugon Cho ◽

Stephen Shemlon ◽

Heesung Jun ◽

Stanley M. Dunn

Keyword(s):

Structural Analysis ◽

Muscle Cell ◽

Electron Micrographs ◽

Relative Size ◽

Automatic Classification ◽

Nerve Fibers ◽

Analysis Algorithm ◽

Structural Networks

Introduction: The analysis and interpretation of electron micrographs of cells and tissues, often requires the accurate extraction of structural networks, which either provide immediate 2D or 3D information, or from which the desired information can be inferred. The images of these structures contain lines and/or curves whose orientation, lengths, and intersections characterize the overall network.Some examples exist of studies that have been done in the analysis of networks of natural structures. In, Sebok and Roemer determine the complexity of nerve structures in an EM formed slide. Here the number of nodes that exist in the image describes how dense nerve fibers are in a particular region of the skin. Hildith proposes a network structural analysis algorithm for the automatic classification of chromosome spreads (type, relative size and orientation).

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