Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Assembly process”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 865 records · Page 48Linked to original sources
PubMed1997

The role of the electrostatic coat in the formation of cholesteric liquid crystal spherulites from alpha-chitin.

The alpha-chitin used in the experiments came from crab shell waste. This was boiled in 3M HC1 to form a colloidal suspension of chitin crystallites. The electrostatic 'cost' surrounding the chitin was then manipulated in two ways. The first was the alteration of the pH of the chitin colloid (Chitin pKa = 6.1). This allowed the charge density on the crystalline rod of chitin to be altered. The second way was to alter the background charge in the environment by adding salt solutions to the colloid. The effect of the treatments was ascertained by measuring the diameter of the spherulites formed in vitro. These spherulites formed via self assembly through a liquid crystalline cholesteric phase. Raising the pH (within limits), resulted in larger spherulites. Raising the background charge also gave larger spherulites (within limits). As such both background charge and charge on the rod can be used to control the self assembly of the cholesteric spherulites. Manipulation of the electrostatic coat of the chitin could be a method of cellular remote control for formation of the helicoid in arthropod cuticle. This would allow the arthropods to set up conditions that aid the self assembly process.

Animals↗
PubMed1998

The role of pH, temperature and nucleation in the formation of cholesteric liquid crystal spherulites from chitin and chitosan.

The alpha chitin and chitosan used in the experiments came from crab shell waste (Sigma). This was treated to form a colloidal suspension of chitin or chitosan crystallites. The electrostatic 'charge coat' surrounding the chitin was then manipulated. This was achieved by alteration of the pH of the chitin or chitosan colloid (Chitin pKa = 6.1). This allowed the charge density on the crystalline rod of chitin or chitosan to be altered. The effect of this treatment was ascertained by measuring the diameter of spherulites formed in vitro. The results were analysed to see if the experimental optimal pH agreed with theoretical approximations. Further investigations were carried out on the initiation of the spherulites and the effect of temperature on spherulite formation. The spherulites form via self assembly through a liquid crystalline cholesteric phase. Manipulation of the electrostatic coat of the chitin could be a method of cellular remote control for formation of the helicoid in arthropod cuticle. This would allow the arthropods to set up conditions that aid the self assembly process.

Chitin↗
PubMed2003

Mutational analysis of conserved amino acids in the T cell receptor alpha-chain transmembrane region: a critical role of leucine 112 and phenylalanine 127 for assembly and surface expression.

Correct assembly of all TCR complex polypeptides is essential for its cell surface expression and function. The transmembrane region of the TCRalpha chain is highly conserved and to gain insight into the structural and functional role of these residues, single amino acid substitutions were introduced and surface expression and signaling ability studied in T hybridoma cells. Introduction of acid residues within the TCRalpha chain transmembrane region were mostly tolerated, indicating that the net charge within this region of the TCR complex is not crucial to either assembly or signaling. However, mutations of leucine 112 or phenylalanine 127 to aspartic acids (L112D or F127D, respectively) resulted in dramatic loss of surface expression and, therefore, their signaling ability. Intracellular flow cytometry showed that the mutant TCRalpha polypeptides were present at levels comparable to wild-type, indicating that the reduced surface expression was not a consequence of impaired protein survival. The defect was characterized by immunoprecipitation and showed that residues L112 and F127 were involved in early interactions with the CD3 complex. A large proportion of the TCRalpha chain mutants L112D and F127D consisted of immature protein, indicative of a problem during early assembly of the TCR. Our findings provide evidence for the involvement of the conserved L112 and F127 residues of the TCRalpha chain transmembrane region in the assembly process of the TCR complex.

Amino Acid Sequence↗
PubMed1999

Purification of E. coli-expressed HIS-tagged hepatitis B core antigen by Ni2+ -chelate affinity chromatography.

Hepatitis B virus is a major cause of human liver disease. In the case of chronic infection the virus can lead to liver cancer and cirrhosis. The virion consists of an outer envelope containing lipids of the endoplasmic reticulum and virally-encoded surface proteins. This lipoprotein shell encloses the nucleocapsid or core antigen (HBcAg), which contains the viral genome. The capsid consists of dimers of a 183-residue protein, which can be divided into an assembly (residues 1-149) and a protamin-like domain (residues 150-183), responsible for polymerization into particles and RNA packaging, respectively. Upon expression of the core gene in bacteria the products are assembled into capsids resembling those of wild type particles. A purification protocol was developed for unpolymerised (dimeric) and polymerized HBcAg by fusion of six histidine residues to a C-terminal deletion mutant of the core protein allowing the isolation of the respective antigens after denaturing Ni2+-chelate affinity chromatography and renaturing dialysis. The possible incorporation of E. coli proteins during the assembly process and the inclusion of nucleic acids can be avoided. The method might be an attractive alternative to common purification protocols of hybrid virus-like particles (VLPs) for vaccine use.

Chromatography, Affinity↗
PubMed2002

The microsporidian polar tube: evidence for a third polar tube protein (PTP3) in Encephalitozoon cuniculi.

The invasion strategy used by microsporidia is primarily related to spore germination. Small differentiated spores of these fungi-related parasites inject their contents into target cells through the lumen of a rapidly extruded polar tube, as a prerequisite to obligate intracellular development. Previous studies in Encephalitozoon species that infect mammals have identified two major antigenic polar tube proteins (PTP1 and PTP2) which are predicted to contribute to the high tensile strength of the polar tube via an assembly process dependent on disulfide linkages. By immunoscreening of a cDNA library, we found that a novel PTP is encoded by a single transcription unit (3990 bp) located on the chromosome XI of E. cuniculi. PTP3 is predicted to be synthesized as a 1256-amino acid precursor with a cleavable signal peptide. The mature protein lacks cysteine residue and its large acidic core is flanked by highly basic N- and C-terminal regions. Immunolocalization data indicated that PTP3 is involved in the sporoblast-to-spore polar tube biogenesis. A transcriptional up-regulation during sporogony is supported by a strong increase in the relative amount of Ecptp mRNAs within host cells sampled at late post-infection times. To begin to explore polar tube-associated protein interactions, spore proteins were extracted in the presence of SDS and dithiothreitol then incubated with a chemical cross-linker (DSP or sulfo-EGS). A large multimeric complex was formed and shown to contain PTP1, PTP2 and PTP3 with a few other proteins. PTP3 is hypothesized to play a role in the control of the polar tube extrusion as part of a specific response to ionic stimuli.

Amino Acid Sequence↗
PubMed1998

The DnaJ family of protein chaperones in Trypanosoma cruzi.

We have molecularly cloned four members of the DnaJ (heat shock protein 40) family of protein chaperones of the protozoan parasite Trypanosoma cruzi--tcj1, tcj2, tcj3 and tcj4. While all the proteins contain defining J domains at their N-termini, only tcj2, tcj3 and tcj4 contain glycine/phenylalanine-rich and zinc finger domains common to many other DnaJ homologues. Furthermore, tcj2 and tcj4 contain C-terminal CaaX motifs, substrates for prenyl modifications, suggesting that they are associated with cellular membranes. tcj1 is a divergent member of the family, containing neither glycine/phenylalanine-rich nor zinc finger domains. All the T. cruzi DnaJ genes are single copy, in contrast to other T. cruzi heat shock genes, which are arranged in multicopy direct tandem arrays. Among the tcj mRNAs, only tcj2 is heat inducible, which may result from posttranscriptional regulation involving a sequence found in the 3' untranslated regions of all heat-inducible T. cruzi mRNAs described to date. Further study of this important family of protein chaperones will aid our understanding of the protein folding and assembly processes in protozoans.

Amino Acid Sequence↗
PubMed1998

Intrinsic protein electric fields: basic non-covalent interactions and relationship to protein-induced Stark effects.

Knowledge of the interactions involving charged, polar and polarizable groups in proteins is fundamental, not only because they are important determinants for gaining insight into biophysical molecular recognition and assembly processes, but also for understanding how the matrix of a protein can be viewed as an electric field capable of inducing Stark perturbations on the spectral properties of biological optical centers. This review describes the essential features of noncovalent interactions in protein systems and discusses the concept of the dielectric constant of a protein in the context of different microscopic and macroscopic modeling approaches. It also provides an account of a specific type of high resolution vibrational and optical Stark spectroscopy attempting to correlate the observed spectral properties of biological optical centers to the intrinsic protein fields induced by the matrix in which they reside.

Models, Chemical↗
PubMed2003

Polymeric nanoparticle composed of fatty acids and poly(ethylene glycol) as a drug carrier.

Diamine-terminated poly(ethylene glycol) (ATPEG) was hydrophobically modified with long-chain fatty acids (FAs) through a coupling reaction using N,N'-dicyclohexyl carbodiimide (DCC). FA-PEG-FA conjugates have different physico-chemical properties according to the chain length of the fatty acid (FA). Synthesized FA-PEG-FA conjugate was confirmed by Fourier transform-infrared (FT-IR). Since FA-PEG-FA conjugates have the amphiphilic characteristics in aqueous solution, polymeric nanoparticles of FA-PEG-FA conjugates were prepared using a simple dialysis method in water. The results of 1H nuclear magnetic resonance (NMR) spectroscopy and fluorescent spectroscopy suggest that the FA-PEG-FA conjugate has a typical core-shell type nanoparticle structure made by a self-assembling process. From the analysis of fluorescence excitation spectra, especially, the critical micelles concentration (CMC) of this conjugate was changed unpredictably, i.e. the critical association concentration (CAC) value was decreased below a FA carbon number of 16 but, above increased a FA carbon number of 16. Transmission electron micrograph readings showed the spherical morphologies of the polymeric nanoparticles. The particle size was continuously decreased until below a FA carbon number of 20, but it was increased above a FA carbon number of 20. Clonazepam (CNZ), as a model drug, was easy to entrap into polymeric nanoparticles of the FA-PEG-FA conjugates. The drug release behavior was changed according to the FA chain length and was mainly diffusion controlled from the core portion.

Drug Carriers↗
PubMed2000

dtrap-1 encodes a novel member of the heat shock super family of proteins and is expressed in derivatives of all three germ layers during Drosophila embryogenesis.

Heat shock proteins (Hsps) comprise a highly conserved superfamily of proteins that are required for stress tolerance in living cells. At physiological conditions, these proteins act as chaperones during protein folding and protein assembly processes (Parsell and Lindquist, 1993. Annu. Rev. Genet. 27, 427-496). Members of the subfamily of Hsp90 proteins were shown to be additionally involved in the presentation and structural modification of components of diverse cellular signal transduction pathways including steroid hormone reception and regulatory kinase activities (Pratt, 1993. J. Biol. Chem. 268, 21455-21458); Cutforth et al., 1994. Cell 77, 1027-1036; van der Straten et al., 1997. EMBO J. 16, 1961-1997; Hunter and Poon, 1997. Trends Cell Biol. 7, 157-161). We have identified a Drosophila gene, called dtrap-1, which encodes a Hsp of a novel subfamily that is related to the Hsp90 family of proteins. During oogenesis dtrap-1 is expressed in nurse cells and its transcripts accumulate in the oocyte. The maternal transcripts remain in the egg but rapidly degrade during early embryogenesis, except in the posterior pole region. Zygotic expression is initiated after the onset of gastrulation showing dynamic patterns of transcripts in the developing mid- and hindgut as well as a subset of mesoderm derivatives.

Animals↗
PubMed1991

A discrete helicoid of collagenous fibrils: the cuticle of deep-sea hydrothermal vent worms (Riftia pachyptila).

The fibrillar organization of the collagenous cuticle of the hydrothermal vent worm Riftia pachyptila is described. Fibrils in the posterior part of the cuticle are organized in a classical orthogonal plywood consisting of successive layers of fibrils: in this case, fibrils are oriented in only two directions which are orthogonal, as for pogonophoran. Our new data on the plume of Riftia pachyptila show a new type of fibrillar arrangement of the cuticle: 1) three sets of fibrils are arranged in an hexagonal pattern; 2) fibrils in successive plies are rotated by 60 degrees, and the organization of the fibrillar network is interpreted as a discrete helicoid when compared to continuously twisted plywoods; 3) a fourth set of fibrils crosses the hexagonally arranged plies and is oriented perpendicular to the surface of the body. X-ray diffraction studies of the cuticular fibrils reveal a triple helix which is characteristic of collagen molecules. Results obtained by differential scanning calorimetry (DSC) show that the denaturation temperature of the molecule is 54.7 degrees C for Riftia; whereas it is 58.9 degrees C for type I collagen measured under the same conditions. We discuss the origin of this plywood with respect to biomechanical constraints, self assembly processes, and compartmentation of the extracellular space. The involvement of the cell membrane in the fibrillogenesis of collagen is also discussed.

Animals↗
PubMed2002

The SMN complex, an assemblyosome of ribonucleoproteins.

Spinal muscular atrophy is a common, often lethal, neurodegenerative disease that results from low levels of, or loss-of-function mutations in, the SMN (survival of motor neurons) protein. SMN oligomerizes and forms a stable complex with five additional proteins: Gemins 2-6. SMN also interacts with several additional proteins referred to as "substrates". Most of these substrates contain a domain enriched in arginine and glycine residues (the RG-rich domain), and are constituents of different ribonucleoprotein complexes. Recent studies revealed that the substrates can be modified by an arginine methyltransferase complex, the methylosome. This forms symmetrical dimethylarginines within the RG-rich domains of the substrates, thereby converting them to high-affinity binders of the SMN complex, and most likely providing regulation of the ribonucleoprotein assembly processes.

Amino Acid Sequence↗
PubMed2001

Microgravimetric DNA sensor based on quartz crystal microbalance: comparison of oligonucleotide immobilization methods and the application in genetic diagnosis.

We report on the study of immobilization DNA probes onto quartz crystal oscillators by self-assembly technique to form variety types of mono- and multi-layered sensing films towards the realization of DNA diagnostic devices. A 18-mer DNA probe complementary to the site of genetic beta-thalassaemia mutations was immobilized on the electrodes of QCM by covalent bonding or electrostatic adsorption on polyelectrolyte films to form mono- or multi-layered sensing films by self-assembled process. Hybridization was induced by exposure of the QCMs immobilized with DNA probe to a test solution containing the target nucleic acid sequences. The kinetics of DNA probe immobilization and hybridization with the fabricated DNA sensors were studied via in-situ frequency changes. The characteristics of QCM sensors containing mono- or multi-layered DNA probe constructed by direct chemical bonding, avidin-biotin interaction or electrostatic adsorption on polyelectrolyte films were compared. Results indicated that the DNA sensing films fabricated by immobilization of biotinylated DNA probe to avidin provide fast sensor response and high hybridization efficiencies. The effects of ionic strength of the buffer solution and the concentration of target nucleic acid used in hybridization were also studied. The fabricated DNA biosensor was used to detect a set of real samples. We conclude that the microgravimetric DNA sensor with its direct detection of amplified products provide a rapid, low cost and convenient diagnostic method for genetic disease.

Biosensing Techniques↗
PubMed2003

Patterning of photosensitive polyimide LB film and its application in the fabrication of biomolecular microphotodiode array.

Ultra thin film of photosensitive polyimide having benzene and sulfonyloxyimide moieties in the main chain was prepared using a Langmuir-Blodgett (LB) technique, and then micro array pattern of the polyimide LB film on a gold substrate was obtained by deep UV lithographic technique. In order to array cytochrome c molecules along the micro-patterned gold substrate, the well-characterized monolayer of cytochrome c was immobilized with a mixed monolayer of 11-mercaptoundecanoic acid (11-MUDA) and decanethiol. The redox activity and electron transfer between cytochrome c molecular center and gold electrode interface for the self-assembled cytochrome c monolayer were investigated by cyclic voltammetry measurement. Biomolecular photodiode consisting of cytochrome c and green fluorescent protein (GFP) onto the patterned gold substrate was fabricated by self-assembly process. The integration and morphology of cytochrome c and GFP were studied from the measurements of atomic force microscopy (AFM) and fluorescence emission. Especially, current-voltage characteristics of the protein multilayers were investigated by scanning tunneling microscopy (STM) and its application in biomolecular photodiode was also examined.

Adsorption↗
PubMed2003

The role of mobile genetic elements in bacterial adaptation to xenobiotic organic compounds.

Retrospective studies clearly indicate that mobile genetic elements (MGEs) play a major role in the in situ spread and even de novo construction of catabolic pathways in bacteria, allowing bacterial communities to rapidly adapt to new xenobiotics. The construction of novel pathways seems to occur by an assembly process that involves horizontal gene transfer: different appropriate genes or gene modules that encode different parts of the novel pathway are recruited from phylogenetically related or distant hosts into one single host. Direct evidence for the importance of catabolic MGEs in bacterial adaptation to xenobiotics stems from observed correlations between catabolic gene transfer and accelerated biodegradation in several habitats and from studies that monitor catabolic MGEs in polluted sites.

Adaptation, Physiological↗
PubMed2000

Theoretical studies of viral capsid proteins.

Recent results in structural biology and increases in computer power have prompted initial theoretical studies on capsids of nonenveloped icosahedral viruses. The macromolecular assembly of 60 to 180 protein copies into a protein shell results in a structure of considerable size for molecular dynamics simulations. Nonetheless, progress has been made in examining these capsid assemblies from molecular dynamics calculations and kinetic models. The goals of these studies are to understand capsid function and structural properties, including quarternary structural stability, effects of antiviral compounds that bind the capsid and the self-assembly process. The insight that can be gained from the detailed information provided by simulations is demonstrated in studies of human rhinovirus; an entropic basis for the antiviral activity of hydrophobic compounds, predicted from calculated compressibility values, has been corroborated by experimental measurements on poliovirus.

Antiviral Agents↗
PubMed1998

Association of the rat liver glucocorticoid receptor with Hsp90 and Hsp70 upon whole body hyperthermic stress.

The influence of whole body hyperthermic stress (41 degrees C, 15 min) on association of the glucocorticoid receptor (GR) with heat shock proteins Hsp90 and Hsp70 was followed in rat liver cytosol during a 24 h period after the stress. Total cytosolic concentration of the GR, Hsp90 and Hsp70 and the amounts of Hsp90 and Hsp70 co-immunopurified with the GR were determined by a quantitative Western blotting using appropriate monoclonal antibodies. A significant decrease in the cytosolic GR level in response to the stress was noticed. The ratio of the amount of the GR to Hsp90 recruited by the GR was found to be unaltered by hyperthermia, in spite of the stress-induced increase in the total Hsp90 concentration in the cytosol. Hsp70 was also found in association with the GR and its 2.5-fold induction by the stress was accompanied by about 3-fold increase in its relative amount that co-immunopurified with the GR. The results suggest that heat stress influences the interaction of the GR with Hsp70 through the mechanisms controlling the untransformed rat liver GR heterocomplexes assembly process.

Animals↗
PubMed1995

The microtubule lattice--20 years on.

In 1974, optical diffraction and image analysis indicated that tubulin dimers in the cylindrically complete A-tubule of flagellar doublet microtubules are arranged with helical symmetry, while those in the incomplete B-tubule associate differently. Recently, electron micrographs of reassembled brain microtubules decorated with kinesin heads have shown that the tubulin dimers there are arranged as in the B-tubule. The lack of symmetry of microtubules assembled in vitro prompts Linda Amos to speculate here that the assembly process in vitro may differ from that occurring in the cell.

Journal Article↗
PubMed2000

Understanding protein folding via free-energy surfaces from theory and experiment.

The ability of protein molecules to fold into their highly structured functional states is one of the most remarkable evolutionary achievements of biology. In recent years, our understanding of the way in which this complex self-assembly process takes place has increased dramatically. Much of the reason for this advance has been the development of energy surfaces (landscapes), which allow the folding reaction to be described and visualized in a meaningful manner. Analysis of these surfaces, derived from the constructive interplay between theory and experiment, has led to the development of a unified mechanism for folding and a recognition of the underlying factors that control the rates and products of the folding process.

Animals↗