Search results for "Structural Biology."

Transmission Electron Microscopy of GroEL, GroES, and the Symmetrical GroEL/ES Complex

1994

Two new 2-D crystal forms of the Escherichia coli chaperone GroEL (cpn60) 2 x 7-mer have been produced using the negative staining-carbon film (NS-CF) technique. These 2-D crystals, which contain the cylindrical GroEL in side-on and end-on orientations, both possess p21 symmetry, with two molecules in the respective unit cells. The crystallographically averaged images correlate well with those obtained by other authors from single particle analysis of GroEL and our own previous crystallographic analysis. 2-D crystallization of the smaller chaperone GroES (cpn10) 7-mer has also been achieved using the NS-CF technique. Crystallographically averaged images of GroES single particle images indic…

Structure of keyhole limpet hemocyanin type 1 (KLH1) at 15 Å resolution by electron cryomicroscopy and angular reconstitution † 1 †This article is de…

1997

Abstract A three-dimensional reconstruction of keyhole limpet hemocyanin type 1 (KLH1) has been obtained using electron cryomicroscopy at liquid helium temperatures and single particle image processing. The use of a high-contrast embedding medium, 1% (w/v) glucose and 2% (w/v) ammonium molybdate (pH 7.0), enables high-resolution electron micrographs to be recorded close to focus, i.e. with excellent transfer of high-resolution information, while maintaining enough image contrast to localise the individual macromolecules in the images. When low-pass filtered to ∼45 A resolution, the new 15 A resolution reconstruction is very similar to the earlier reconstructions of gastropodan hemocyanins o…

Review of the shore-fly genus Oedenopiforma (Diptera: Ephydridae)

2012

The shore-fly genus Oedenopiforma Cogan is reviewed, including new information on Oedenopiforma argentea (distribution), Oedenopiforma javana (comb. nov. and O. orientalis syn. nov.), and Oedenopiforma vockerothi (sp. nov. from United Arab Emirates). Structures of the male terminalia of these three species, a diagnosis of the genus, and a key to the included genera of Dryxini and species of Oedenopiforma are presented. Resume´—Nous revisons le genre Oedenopiforma Cogan chez les ephydrides et apportons de nouvelles informations sur Oedenopiforma argentea (repartition), Oedenopiforma javana (nouvelle combinaison et O. orientalis nouvelle synonymie )e tOedenopiforma vockerothi (nouvelle espece…

Conformational dynamics of full-length inducible human Hsp70 derived from microsecond molecular dynamics simulations in explicit solvent

2013

Human 70 kDa heat shock protein (hHsp70) is an ATP-dependent chaperone and is currently an important target for developing new drugs in cancer therapy. Knowledge of the conformations of hHsp70 is central to understand the interactions between its nucleotide-binding domain (NBD) and substrate-binding domain (SBD) and is a prerequisite to design inhibitors. The conformations of ADP-bound (or nucleotide-free) hHsp70 and ATP-bound hHsp70 was investigated by using unbiased all-atom molecular dynamics (MD) simulations of homology models of hHsp70 in explicit solvent on a timescale of .5 and 2.7 μs, respectively. The conformational heterogeneity of hHsp70 was analyzed by computing effective free-e…

Production, crystallization and preliminary X-ray analysis of the human integrin alpha1 I domain.

1999

Integrin α1β1 is one of the main collagen receptors in many cell types. A fast large-scale production, purification and crystallization method for the integrin α1 I domain is reported here. The α1 I domain was crystallized using the vapour-diffusion method with a reservoir solution containing a mixture of PEG 4000, sodium acetate, glycerol and Tris–HCl buffer. The crystals beong to the C2 space group, with unit-cell parameters a = 74.5, b = 81.9, c = 37.3 Å, α = γ = 90.0, β = 90.8°. The crystals diffract to 2.0 Å and a 94.2% complete data set to 2.2 Å has been collected from a single crystal with an R merge of 5.8%.

Identification of highly conserved genes: SNZ and SNO in the marine sponge Suberites domuncula: their gene structure and promoter activity in mammali…

2001

Abstract Recently, we reported that cells from the sponge Suberites domuncula respond to ethylene with an increase in intracellular Ca 2+ level [Ca 2+ ] i , and with an upregulation of the expression of (at least) two genes, a Ca 2+ /calmodulin-dependent protein kinase and the potential ethylene-responsive gene, termed SDSNZERR (A. Krasko, H.C. Schroder, S. Perovic, R. Steffen, M. Kruse, W. Reichert, I.M. Muller, W.E.G. Muller, J. Biol. Chem. 274 (1999)). Here, we describe for the first time that also mammalian (3T3) cells respond to ethylene, generated by ethephon, with an immediate and transient, strong increase in [Ca 2+ ] i . Next, the promoter for the sponge SDSNZERR gene was isolated …

Novel structural insights into F-actin-binding and novel functions of calponin homology domains.

2008

Tandem calponin homology (CH) domains are well-known actin filaments (F-actin) binding motifs. There has been a continuous debate about the details of CH domain-actin interaction, mainly because atomic level structures of F-actin are not available. A recent electron microscopy study has considerably advanced our structural understanding of CH domain:F-actin complex. On the contrary, it has recently also been shown that CH domains can bind other macromolecular systems: two CH domains from separate polypeptides Ncd80, Nuf2 can form a microtubule-binding site, as well as tandem CH domains in the EB1 dimer, while the single C-terminal CH domain of alpha-parvin has been observed to bind to a alp…

Evolutionary history and diversity of arthropod hemocyanins

2004

Hemocyanins are copper-containing, multi-subunit proteins that transport oxygen in the hemolymph of many molluscs and arthropods [Markl and Decher, Adv. Comp. Environ. Physiol. 13 (1992) 325; van Holde et al., J. Biol. Chem. 276 (2001) 15563]. Arthropod hemocyanins originated more than 550 million years ago from oxygen-consuming phenoloxidases. Hemocyanins are present in various Onychophora, Chelicerata, Myriapoda, Crustacea, and Hexapoda, but subunit evolution differs striking in these arthropod subphyla. Hemocyanins also gave rise to non-respiratory proteins (crustacean pseudo-hemocyanins, insect hexamerins, and hexamerin receptors), which most likely have storage functions.

Solubilization of an oligomycin-sensitive ATPase complex fromRhodospirillum rubrumchromatophores and its inhibition by various antibiotics

1978

Immunoelectron Microscopy of Hemocyanin from the Keyhole Limpet (Megathura crenulata): A Parallel Subunit Model

1993

Abstract Immunoelectron microscopy has been performed using negatively stained immune complexes of keyhole limpet hemocyanin (KLH) subunit 2 di- and multidecamers with domain-specific monoclonal antibodies. One antibody (KLH2 a macr 1) links the hemocyanin molecules in a side-to-side pattern, whereas the other antibody (KLH2 fg macr 1) links the molecules end-to-end. From existing knowledge of the domain sequence of KLH subunit 2, these data provide support for a parallel arrangement of subunits within each decamer. Ten N-terminal a macr: domains are then present at the noncollar region of each decamer with 10 C-terminal g macr domains at the collar region. The immunonegative staining data …