Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Immunolocalization of lamins and nuclear pore complex proteins by atomic force microscopy.
Schneider S
,
Folprecht G
,
Krohne G
,
Oberleithner H
.
???displayArticle.abstract???
The nuclear envelope functions as a selective barrier separating the nuclear from the cytosolic compartment. Nuclear pore complexes (NPCs) mediate nuclear import and export of macromolecules and, therefore, are potential regulators of gene expression. In this study we applied atomic force microscopy (AFM) to visualize the three dimensional (3D) structure of individual NPCs in the absence and presence of two different antibodies, one directed against a pore protein (gp62) and another directed against Xenopus lamin LIII, a component of the nuclear lamina, a filament meshwork localized on the nucleoplasmic side of the nuclear envelope (NE) adjacent to and interacting with NPCs. Using 12-nm gold-labelled secondary antibodies and transmission electron microscopy we could clearly localize the primary single anti-gp62 antibody on NPCs and the primary single anti-LIII antibody between NPCs. Using AFM, the secondary antibodies against anti-gp62 could be detected as particles 7 nm in height on the nucleoplasmic face of NPCs. The secondary antibodies against anti-LIII could be clearly identified between NPCs. The secondary antibodies, attached to a 12-nm colloidal gold particle and visualized on glass, revealed similar shapes and heights as found on NEs. According to the 3D images, the volume of a single gold particle conjugated with secondary antibodies was 10203 nm3. This volume is equivalent to the volume of 38 IgG molecules associated with one individual gold particle. A similar volume of 11987 nm3 was calculated from a model assuming that the 150-kDa IgG molecules perfectly cover the spherical gold particle. We conclude that AFM can be used for identifying antibodies or other macromolecules associated with biomembranes.
Aaronson,
Isolation of nuclear pore complexes in association with a lamina.
1975, Pubmed
Aaronson,
Isolation of nuclear pore complexes in association with a lamina.
1975,
Pubmed
Aebi,
The nuclear lamina is a meshwork of intermediate-type filaments.
,
Pubmed
,
Xenbase
Akey,
Interactions and structure of the nuclear pore complex revealed by cryo-electron microscopy.
1989,
Pubmed
,
Xenbase
Akey,
Architecture of the Xenopus nuclear pore complex revealed by three-dimensional cryo-electron microscopy.
1993,
Pubmed
,
Xenbase
Benavente,
Cell type-specific expression of nuclear lamina proteins during development of Xenopus laevis.
1985,
Pubmed
,
Xenbase
Binnig,
Atomic force microscope.
1986,
Pubmed
Cordes,
Intranuclear filaments containing a nuclear pore complex protein.
1993,
Pubmed
,
Xenbase
Cordes,
Nuclear pore complex glycoprotein p62 of Xenopus laevis and mouse: cDNA cloning and identification of its glycosylated region.
1991,
Pubmed
,
Xenbase
Edstrom,
Direct visualization of phosphorylase-phosphorylase kinase complexes by scanning tunneling and atomic force microscopy.
1990,
Pubmed
Foisner,
Integral membrane proteins of the nuclear envelope interact with lamins and chromosomes, and binding is modulated by mitotic phosphorylation.
1993,
Pubmed
Franke,
Structure, biochemistry, and functions of the nuclear envelope.
1974,
Pubmed
Franke,
The nuclear envelope and the architecture of the nuclear periphery.
1981,
Pubmed
Gerace,
Molecular trafficking across the nuclear pore complex.
1992,
Pubmed
Gieffers,
In vitro reconstitution of recombinant lamin A and a lamin A mutant lacking the carboxy-terminal tail.
1991,
Pubmed
,
Xenbase
Goldberg,
High resolution scanning electron microscopy of the nuclear envelope: demonstration of a new, regular, fibrous lattice attached to the baskets of the nucleoplasmic face of the nuclear pores.
1992,
Pubmed
,
Xenbase
Heitlinger,
The role of the head and tail domain in lamin structure and assembly: analysis of bacterially expressed chicken lamin A and truncated B2 lamins.
1992,
Pubmed
Henderson,
Actin filament dynamics in living glial cells imaged by atomic force microscopy.
1992,
Pubmed
,
Xenbase
Hinshaw,
Architecture and design of the nuclear pore complex.
1992,
Pubmed
,
Xenbase
Hoh,
Atomic force microscopy for high-resolution imaging in cell biology.
1992,
Pubmed
Hoh,
Atomic force microscopy and dissection of gap junctions.
1991,
Pubmed
Jarnik,
Toward a more complete 3-D structure of the nuclear pore complex.
1991,
Pubmed
,
Xenbase
McKeon,
Homologies in both primary and secondary structure between nuclear envelope and intermediate filament proteins.
,
Pubmed
Moir,
Expression in Escherichia coli of human lamins A and C: influence of head and tail domains on assembly properties and paracrystal formation.
1991,
Pubmed
Nigg,
Assembly-disassembly of the nuclear lamina.
1992,
Pubmed
Oberleithner,
Imaging the lamellipodium of migrating epithelial cells in vivo by atomic force microscopy.
1993,
Pubmed
Oberleithner,
Imaging nuclear pores of aldosterone-sensitive kidney cells by atomic force microscopy.
1994,
Pubmed
Panté,
The nuclear pore complex.
1993,
Pubmed
Radmacher,
Direct observation of enzyme activity with the atomic force microscope.
1994,
Pubmed
Radmacher,
From molecules to cells: imaging soft samples with the atomic force microscope.
1992,
Pubmed
Reichelt,
Correlation between structure and mass distribution of the nuclear pore complex and of distinct pore complex components.
1990,
Pubmed
,
Xenbase
Riemer,
A nuclear lamin of the nematode Caenorhabditis elegans with unusual structural features; cDNA cloning and gene organization.
1993,
Pubmed
Scheer,
Experimental disintegration of the nuclear envelope. Evidence for pore-connecting fibrils.
1976,
Pubmed
,
Xenbase
Simos,
The inner nuclear membrane protein p58 associates in vivo with a p58 kinase and the nuclear lamins.
1992,
Pubmed
Unwin,
A large particle associated with the perimeter of the nuclear pore complex.
1982,
Pubmed
,
Xenbase
Wilken,
Nup180, a novel nuclear pore complex protein localizing to the cytoplasmic ring and associated fibrils.
1993,
Pubmed
,
Xenbase
