Journal of Structural Biology
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- Sciencedirect from 1990, Ingenta, Gale, Rcgp, Proquest, and CSA
2D and 3D crystallization of a bacterial homologue of human vitamin C membrane transport proteins.
We cloned, overexpressed, purified, functionally characterized, and 2D- and 3D-crystallized a bacterial protein (UraDp) with 29% of amino acid sequence identity to the human sodium-dependent vitamin C transporter 1 (SVCT1). Ligand-binding experiments by scintillation proximity assay revealed that ur...
Mapping the road to recovery: the ClpB/Hsp104 molecular chaperone.
We focus on one family of Hsp100s, the homologous ClpB and Hsp104 molecular chaperones that convey thermotolerance by resolubilising and rescuing proteins from aggregates. We explore how the nucleotide binding and hydrolysis properties at the twelve nucleotide-binding domains of these hexameric ring...
Large nuclear RNP particles—the nuclear pre-mRNA processing machine
We show that the cap-binding proteins (CBPs), CBP20 and CBP80, are associated with the lnRNP particles, as well as components of the 3′-end-processing activity. These results, together with our previous demonstration of the association of splicing factors and A-to-I editing enzymes with lnRNP part...
The distribution of RNA polymerase II largest subunit (RPB1) in the Xenopus germinal vesicle
We present evidence that this stain is due to cross-reaction with one or more SR proteins. We show that constructs consisting of 15–17 CTD heptapeptide repeats fused to glutathione-S-transferase are targeted rapidly and specifically to CBs but not to B-snurposomes after injection into the nucleus....
On the formation of amplified nucleoli during early Xenopus oogenesis
We have studied nucleolar development in early previtellogenic Xenopus oocytes. By using light and electron microscopic immunocytochemistry, in situ hybridization, and the TdT-method for detection of DNA, we have identified three separate structural entities which are related to the three components...
Single molecule imaging of supported planar lipid bilayer—reconstituted human insulin receptors by in situ scanning probe microscopy
We report here the results of an in situ study by high-resolution scanning probe microscopy of the full-length insulin receptor reconstituted within supported planar lipid bilayers. Our preliminary studies confirm that (1) the intact receptor can be reconstituted constitutively within a lipid vesicl...
Use of frozen-hydrated axonemes to assess imaging parameters and resolution limits in cryoelectron tomography
We have obtained tomographic reconstructions of frozen-hydrated sea urchin axonemes with 8–10-nm resolution, as assessed by detection of characteristic components including doublet microtubules, radial spokes, central sheath projections, and outer dynein arms. We did not detect the inner dynein ar...
Removing high contrast artifacts via digital inpainting in cryo-electron tomography: an application of compressed sensing.
We propose digital inpainting via compressed sensing (CS) as a new method to achieve this goal. We show that cryo-ET projections are sparse in the discrete cosine transform (DCT) domain, and, by finding the sparsest DCT domain decompositions given uncorrupted pixels, we can fill in the missing pixel...
Comparative structural analysis of eukaryotic flagella and cilia from Chlamydomonas, Tetrahymena, and sea urchins.
We here compare the axonemal structure of three different organisms with widely divergent bending-motions by electron cryo-tomography. We reconstruct the 3D structure of the axoneme of Tetrahymena cilia, and compare it with the axoneme of the flagellum of sea urchin sperm, as well as with the axonem...
X-ray mosaic nanotomography of large microorganisms
We exploit the assets of the hard X-ray sub-micrometer imaging and extend the standard approach by widening the effective field of view to match the size of the sample. We show that global tomography of biological systems exceeding several times the field of view is feasible also at the nanoscale wi...