SEARCH · PubMed Health

Results for “Protein aggregation”

Explore indexed PubMed citations for clinical trials, systematic reviews and public health research. Read source abstracts and follow each citation to its original PubMed record.

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

At least 505 records · Page 28Linked to original sources

PubMed2002

Peptide aggregation in neurodegenerative disease.

In the not-so-distant past, insoluble aggregated protein was considered as uninteresting and bothersome as yesterday's trash. More recently, protein aggregates have enjoyed considerable scientific interest, as it has become clear that these aggregates play key roles in many diseases. In this review, we focus attention on three polypeptides: beta-amyloid, prion, and huntingtin, which are linked to three feared neurodegenerative diseases: Alzheimer's, "mad cow," and Huntington's disease, respectively. These proteins lack any significant primary sequence homology, yet their aggregates possess very similar features, specifically, high beta-sheet content, fibrillar morphology, relative insolubility, and protease resistance. Because the aggregates are noncrystalline, secrets of their structure at nanometer resolution are only slowly yielding to X-ray diffraction, solid-state NMR, and other techniques. Besides structure, the aggregates may possess similar pathways of assembly. Two alternative assembly pathways have been proposed: the nucleation-elongation and the template-assisted mode. These two modes may be complementary, not mutually exclusive. Strategies for interfering with aggregation, which may provide novel therapeutic approaches, are under development. The structural similarities between protein aggregates of dissimilar origin suggest that therapeutic strategies successful against one disease may have broad utility in others.

Amino Acid Sequence↗

PubMed1998

High hydrostatic pressure can reverse aggregation of protein folding intermediates and facilitate acquisition of native structure.

The present work demonstrates that high hydrostatic pressure can increase protein folding by reducing nonspecific aggregation. Protein aggregation is one of the main side reactions that competes with protein folding, and it typically results from interactions among partially folded intermediates. It is known that oligomeric proteins can be dissociated by the application of high hydrostatic pressure. Since protein aggregates can be described as nonspecific protein oligomers, it can be predicted that they can be completely or partially dissociated by pressure. The enzyme rhodanese is prone to slow aggregation in 3.9 M urea, and it is widely used as a model for the folding of a protein which readily aggregates. In the present study, it was demonstrated that this aggregation process could be completely reversed under high hydrostatic pressure. Release of the pressure led to renewed protein aggregation. In addition, it was demonstrated that refolding of urea-denatured rhodanese at 2 kbar pressure led to an increased yield of the native enzyme. The final recovery was increased up to approximately 25% in contrast to approximately 5% recovery observed under ambient pressure. The recovery can be further increased in the presence of 4 M glycerol, where 56% of the protein was recovered by treatment with high pressure. These observations suggest that some protein aggregation can be limited without the use of chemical additives, and they show that the pressures needed to maintain solubility are considerably less than those typically required for dissociation of specific oligomers and unfolding of polypeptide chains.

Buffers↗

PubMed2001

Chemical chaperones regulate molecular chaperones in vitro and in cells under combined salt and heat stresses.

Salt and heat stresses, which are often combined in nature, induce complementing defense mechanisms. Organisms adapt to high external salinity by accumulating small organic compounds known as osmolytes, which equilibrate cellular osmotic pressure. Osmolytes can also act as "chemical chaperones" by increasing the stability of native proteins and assisting refolding of unfolded polypeptides. Adaptation to heat stress depends on the expression of heat-shock proteins, many of which are molecular chaperones, that prevent protein aggregation, disassemble protein aggregates, and assist protein refolding. We show here that Escherichia coli cells preadapted to high salinity contain increased levels of glycine betaine that prevent protein aggregation under thermal stress. After heat shock, the aggregated proteins, which escaped protection, were disaggregated in salt-adapted cells as efficiently as in low salt. Here we address the effects of four common osmolytes on chaperone activity in vitro. Systematic dose responses of glycine betaine, glycerol, proline, and trehalose revealed a regulatory effect on the folding activities of individual and combinations of chaperones GroEL, DnaK, and ClpB. With the exception of trehalose, low physiological concentrations of proline, glycerol, and especially glycine betaine activated the molecular chaperones, likely by assisting local folding in chaperone-bound polypeptides and stabilizing the native end product of the reaction. High osmolyte concentrations, especially trehalose, strongly inhibited DnaK-dependent chaperone networks, such as DnaK+GroEL and DnaK+ClpB, likely because high viscosity affects dynamic interactions between chaperones and folding substrates and stabilizes protein aggregates. Thus, during combined salt and heat stresses, cells can specifically control protein stability and chaperone-mediated disaggregation and refolding by modulating the intracellular levels of different osmolytes.

Adaptation, Biological↗

PubMed1975

Cell surface lipids and adhesion. I. The effects of lysophosphatidyl compounds, phospholipase A2 and aggregation-inhibiting protein.

Aggregation-inhibiting protein (AIP: Curtis & Greaves, 1965), which diminishes the adhesiveness of cells, particularly at low temperatures, is identified in the present paper as phospholipase A2 (EC. 3.1.1.4). Our reasons for this identification are because phospholipase activity parallels AIP activity on cell adhesion, and because various inhibitors and sera act in a parallel manner on adhesion in the presence of AIP or phospholipase. We suggest that the enzyme acts on adhesion by producing lysolecithin and other lysolipids in the plasmalemma. Addition of lysolipids diminishes cell adhesion in a manner similar to phospholipase A. Incubation of cells in Hanks' medium at 37 degrees C has a parallel effect. Conditions which would be expected to stimulate reacylation of lysolipids in the plasmalemma, i.e. incubation of cells in the external presence of CoA, ATP and a fatty acid, lead to a recovery or maintenance of adhesion after or during Hanks' incubation at 37 degrees C. All these results suggest that lipid components of the cell, probably in the plasmalemma, are of importance in adhesion. The results are discussed in relation to the long-standing controversy about the effects of low temperatures and trypsinization on cell adhesion, for phospholipase treatment of cells affects adhesion in a manner similar to trypsinization.

Acylation↗

PubMed1998

Folding and aggregation of designed proteins.

Protein aggregation is studied by following the simultaneous folding of two designed identical 20-letter amino acid chains within the framework of a lattice model and using Monte Carlo simulations. It is found that protein aggregation is determined by elementary structures (partially folded intermediates) controlled by local contacts among some of the most strongly interacting amino acids and formed at an early stage in the folding process.

Amino Acid Sequence↗

PubMed1994

[Fragmentation of Ehrlich ascites carcinoma DNA during influences causing aggregation of cytoskeletal proteins].

Upon exposures of Ehrlich ascites carcinoma cells to heat shock (44 degrees, 1 hr), oxidative stress or energy deprivation, their DNA undergoes fragmentation (35-45% after 5 hrs of incubation) which is considered as a hallmark of apoptosis. Prior to DNA fragmentation the cells exhibited blebbing (55-90% after 1 hr), thus being suggestive of cytoskeletal damage and a 1.5-2-fold increase in the Triton-insoluble protein concentration (protein aggregation) after 3 hrs. Rapid cell death (75% after 4 hrs) occurred only under oxidative stress. Electrophoresis of the Triton-insoluble protein fraction revealed that the common feature of all stress exposures used in this study was a dramatic increase in the aggregation of cytoskeletal proteins--actin and the 57 kDa protein. No dependence of DNA fragmentation on intracellular Ca2+ increase was found. Both DNA fragmentation and protein aggregation were suppressed by glucose, whereas Zn2+, an endonuclease inhibitor, suppressed only DNA fragmentation without any effect on protein aggregation. It is suggested that cytoskeletal damage may trigger tumor cell apoptosis.

Animals↗

PubMed1999

Lattice simulations of aggregation funnels for protein folding.

A computer model of protein aggregation competing with productive folding is proposed. Our model adapts techniques from lattice Monte Carlo studies of protein folding to the problem of aggregation. However, rather than starting with a single string of residues, we allow independently folding strings to undergo collisions and consider their interactions in different orientations. We first present some background into the nature and significance of protein aggregation and the use of lattice Monte Carlo simulations in understanding other aspects of protein folding. The results of a series of simulation experiments involving simple versions of the model illustrate the importance of considering aggregation in simulations of protein folding and provide some preliminary understanding of the characteristics of the model. Finally, we discuss the value of the model in general and of our particular design decisions and experiments. We conclude that computer simulation techniques developed to study protein folding can provide insights into protein aggregation, and that a better understanding of aggregation may in turn provide new insights into and constraints on the more general protein folding problem.

Algorithms↗

PubMed2004

Genome-wide RNA interference screen identifies previously undescribed regulators of polyglutamine aggregation.

Protein misfolding and the formation of aggregates are increasingly recognized components of the pathology of human genetic disease and hallmarks of many neurodegenerative disorders. As exemplified by polyglutamine diseases, the propensity for protein misfolding is associated with the length of polyglutamine expansions and age-dependent changes in protein-folding homeostasis, suggesting a critical role for a protein homeostatic buffer. To identify the complement of protein factors that protects cells against the formation of protein aggregates, we tested transgenic Caenorhabditis elegans strains expressing polyglutamine expansion yellow fluorescent protein fusion proteins at the threshold length associated with the age-dependent appearance of protein aggregation. We used genome-wide RNA interference to identify genes that, when suppressed, resulted in the premature appearance of protein aggregates. Our screen identified 186 genes corresponding to five principal classes of polyglutamine regulators: genes involved in RNA metabolism, protein synthesis, protein folding, and protein degradation; and those involved in protein trafficking. We propose that each of these classes represents a molecular machine collectively comprising the protein homeostatic buffer that responds to the expression of damaged proteins to prevent their misfolding and aggregation.

Animals↗

PubMed2000

Formation of high molecular weight complexes of mutant Cu, Zn-superoxide dismutase in a mouse model for familial amyotrophic lateral sclerosis.

Deposition of aggregated protein into neurofilament-rich cytoplasmic inclusion bodies is a common cytopathological feature of neurodegenerative disease. How-or indeed whether-protein aggregation and inclusion body formation cause neurotoxicity are presently unknown. Here, we show that the capacity of superoxide dismutase (SOD) to aggregate into biochemically distinct, high molecular weight, insoluble protein complexes (IPCs) is a gain of function associated with mutations linked to autosomal dominant familial amyotrophic lateral sclerosis. SOD IPCs are detectable in spinal cord extracts from transgenic mice expressing mutant SOD several months before inclusion bodies and motor neuron pathology are apparent. Sequestration of mutant SOD into cytoplasmic inclusion bodies resembling aggresomes requires retrograde transport on microtubules. These data indicate that aggregation and inclusion body formation are mechanistically and temporally distinct processes.

Animals↗

PubMed2002

Aggregate formation and the structure of the aggregates of disulfide-reduced proteins.

Aggregate formation and the structure of the aggregates of disulfide-reduced proteins were investigated using alpha-lactalbumin and lysozyme as model proteins. First, reducing conditions were adjusted so that only one of the four disulfide bonds present in each native protein was cleaved. These three-disulfide (3SS) proteins are known to adopt almost native conformations, yet formed precipitates with a basic peptide, lactoferricin, and heparin and heparin fragment, respectively, at concentrations at which native proteins mixed with these compounds remained clear. The 3SS-lysozyme also formed precipitates in the absence of these ligands. Thus, subtle structural changes could lead to aggregation. Electron microscopy revealed fibrillar structures in the aggregates of extensively reduced proteins in the absence of ligands but not in their presence, which shows that the reduction of disulfide bonds suffices for fibril formation and that ligands inhibit fibril formation.

Animals↗

PubMed2006

Small molecule inhibitors of lysozyme amyloid aggregation.

Protein amyloid aggregation is associated with a number of important human pathologies, but the precise mechanisms underlying the toxicity of amyloid aggregates are still incompletely understood. In this context, drugs capable of blocking or interfering with the aggregation of amyloidogenic proteins should be considered in strategies aimed at the development of novel therapeutic agents. Human lysozyme variants have been shown to form massive amyloid deposits in the livers and kidneys of individuals affected by hereditary systemic amyloidosis. Currently, there are no clinical treatments available to prevent or reverse formation of such amyloid deposits. We have recently described a number of di- and trisubstituted aromatic compounds that block the formation of soluble oligomers and amyloid fibrils of the beta-amyloid peptide (Abeta) and protect hippocampal neurons in culture from Abeta-induced toxicity. Here, we show that some of those compounds inhibit the formation and disrupt preformed amyloid fibrils from both human and hen egg white lysozyme. These results suggest that these small molecule compounds may serve as prototypes for the development of drugs for the prevention or treatment of different types of amyloidoses.

Aminophenols↗

PubMed2005

Conversion of green fluorescent protein into a toxic, aggregation-prone protein by C-terminal addition of a short peptide.

A non-natural 16-residue "degron" peptide has been reported to convey proteasome-dependent degradation when fused to proteins expressed in yeast (Gilon, T., Chomsky, O., and Kulka, R. (2000) Mol. Cell. Biol. 20, 7214-7219) or when fused to green fluorescent protein (GFP) and expressed in mammalian cells (Bence, N. F., Sampat, R. M., and Kopito, R. R. (2001) Science 292, 1552-1555). We find that expression of the GFP::degron in Caenorhabditis elegans muscle or neurons results in the formation of stable perinuclear deposits. Similar perinuclear deposition of GFP::degron was also observed upon transfection of primary rat hippocampal neurons or mouse Neuro2A cells. The generality of this observation was supported by transfection of HEK 293 cells with both GFP::degron and DsRed(monomer)::degron constructs. GFP::degron expressed in C. elegans is less soluble than unmodified GFP and induces the small chaperone protein HSP-16, which co-localizes and co-immunoprecipitates with GFP::degron deposits. Induction of GFP::degron in C. elegans muscle leads to rapid paralysis, demonstrating the in vivo toxicity of this aggregating variant. This paralysis is suppressed by co-expression of HSP-16, which dramatically alters the subcellular distribution of GFP::degron. Our results suggest that in C. elegans, and perhaps in mammalian cells, the degron peptide is not a specific proteasome-targeting signal but acts instead by altering GFP secondary or tertiary structure, resulting in an aggregation-prone form recognized by the chaperone system. This altered form of GFP can form toxic aggregates if its expression level exceeds the capacity of chaperone-based degradation pathways. GFP::degron may serve as an instructive "generic" aggregating control protein for studies of disease-associated aggregating proteins, such as huntingtin, alpha-synuclein, and the beta-amyloid peptide.

Amino Acid Sequence↗

PubMed2006

Reducing haziness in white wine by overexpression of Saccharomyces cerevisiae genes YOL155c and YDR055w.

Grape proteins aggregate in white wine to form haze. A novel method to prevent haze in wine is the use of haze protective factors (Hpfs), specific mannoproteins from Saccharomyces cerevisiae, which reduce the particle size of the aggregated proteins. Hpf1p was isolated from white wine and Hpf2p from a synthetic grape juice fermentation. Putative structural genes, YOL155c and YDR055w, for these proteins were identified from partial amino acid sequences of Hpf1p and Hpf2p, respectively. YOL155c also has a homologue, YIL169c, in S. cerevisiae. Comparison of the partial amino acid sequence of deglycosylated-Hpf2p with the deduced protein sequence of YDR055w, confirmed five of the 15 potential N-linked glycosylation sites in this sequence were occupied. Methylation analysis of the carbohydrate moieties of Hpf2p indicated that this protein contained both N- and O-linked mannose chains. Material from fermentation supernatant of deletion strains had significantly less activity than the wild type. Moreover, YOL155c and YIL169c overexpressing strains and a strain overexpressing 6xHis-tagged Hpf2p produced greater haze protective activity than the wild type strains. A storage trial demonstrated the short to midterm stability of 6xHis-tagged Hpf2p in wine.

Amino Acid Sequence↗

PubMedDate not supplied

Protein misfolding in Alzheimer's and Parkinson's disease: genetics and molecular mechanisms.

The accumulation of altered proteins is a common pathogenic mechanism in several neurodegenerative disorders. A causal role of protein aggregation was originally proposed in Alzheimer's disease (AD) where extracellular deposition of beta-amyloid (Abeta) is the main neuropathological feature. It is now believed that intracellular deposition of aggregated proteins may be relevant in Parkinson's disease (PD), amyotrophic lateral sclerosis and polyglutamine disorders. An impairment of ubiquitin-proteasome system (UPS) appears directly involved in these disorders. We reviewed the results on the role of protein misfolding in AD and PD and the influence of mutations associated with these diseases on the expression of amyloidogenic proteins. Results of genetic screening of familial cases of AD and PD are summarized. In the familial AD population (70 subjects) we found several mutations of the presenilin 1 (PS1) gene with a frequency of 12.8% and one mutation in the gene encoding the protein precursor of amyloid (APP) (1.4%). One mutation of Parkin in the homozygous form and two in the heterozygous form were identified in our PD population. We also reported data obtained with synthetic peptides and other experimental models, for evaluation of the pathogenic role of mutations in terms of protein misfolding.

Aged↗

PubMed2003

A novel approach of cell-free peptide synthesis and SNPs detection using nano-circular single-stranded DNA.

Development of peptide/protein synthesis is very important for investigating the functions of peptides and proteins. The advantages of cell-free synthesis, which has been extensively developed, include the ability to synthesize toxic proteins, aggregated proteins and proteins containing unnatural amino acids. However, this promising approach does not have a high reaction yield. In this study, we examine whether the rolling synchronization system, which produces long RNAs with a repeated sequence encoded by very small circular ssDNAs without the promoter sequence for RNA polymerase, can be used for cell-free peptide synthesis to improve the reaction yield. As a result, we have found that rolling synchronization is useful for large-scale cell-free peptide/protein synthesis.

Base Sequence↗

PubMed1983

Role of DNA and bacteriophage in Campylobacter auto-agglutination.

Auto-agglutinated and non-agglutinated cells of Campylobacter jejuni and C. coli were examined by transmission electronmicroscopy in phosphotungstate negative stain. Agglutination was induced by three factors (1) extracellular DNA, (2) an aggregated protein, probably a bacteriophage precursor, and (3) free phage-tail sheaths. Auto-agglutinated cells were often "leaky," with a mantle of adhering DNA. About 80% of the auto-agglutinated cells could be resuspended after treatment with DNAase. Flagella were loosely embedded in protein aggregates, especially in phage-infected cultures. They were clumped in a side-by-side arrangement by free phage-tail sheaths. These findings suggest that auto-agglutination could be minimised in suspensions of organisms intended for use in agglutination tests by harvesting early logarithmic-phase cells containing no more than a low phage population. The most common C. jejuni phage had a contractile tail, a head diameter of 60-70 nm, and an overall length of 180-210 nm. A phage isolated from C. jejuni strain 1590 was morphologically identical with C. coli phage.

Agglutination↗

PubMed1996

Analytical criteria for predicting apparent digestibility of soybean protein in preruminant calves.

A series of experiments on the use of soybean as a protein source in milk replacers for veal calves was undertaken to determine the relationships between the physicochemical and antinutritional properties and apparent digestibilities of nine soybean products. Soybean provided between 58 and 71% of dietary CP, and skim milk or whey powder provided the remainder. Soybean products were analyzed for CP, native protein, aggregated protein, carbohydrate-linked protein, peptides, antitryptic activity, and immunoreactivity of lectin, glycinin, alpha-conglycinin, and beta-conglycinin. The apparent digestibility of N calculated for soybean protein varied between 59 and 84%. Simple linear correlations were significant between apparent digestibility of soybean N and concentrations of native protein, antitryptic activity, glycinin, alpha-conglycinin, and beta-conglycinin. However, only variation in antitryptic activity, alpha-conglycinin, and beta-conglycinin contributed significantly to an explanation for the variation in apparent digestibility of soybean N in a multiple linear equation. Under our experimental conditions, beta-conglycinin was the best predictor of digestibility of soybean N. Antitryptic activity became the best predictor when soybean products had no detectable beta-conglycinin.

Animal Nutritional Physiological Phenomena↗

PubMed1995

Spontaneous overexpression of heat-shock proteins in Ehrlich ascites carcinoma cells during in vivo growth.

Ehrlich carcinoma (EC) cells isolated from mice at different phases of ascites growth were exposed to hyperthermia (44 degrees C), or oxidative stress (hydrogen peroxide or vikasol), or ATP depletion induced by rotenone. These exposures caused protein aggregation and rapid necrotic death in exponentially growing EC cells. On the contrary, the same cell culture at stationary phase of growth became considerably more resistant to all the above cytotoxic treatments, and the level of aggregated protein was significantly lower in stressed stationary EC cells than that in exponential ones. Comparative immunoblotting has revealed the unexpected expression of inducible 70 kDa heat-shock protein form (HSP68), as well as accumulation of HSP27 and HSP90 in the thermo- and drug-resistant stationary EC cells. It is suggested that the in vivo occurring HSP overexpression in stationary EC cells is an adaptive modulation of the tumor cell phenotype to maintain the viability of ascites EC cells under chronic deficiency of oxygen and nutrients.

Animals↗