Increasing mechanical strength of gelatin hydrogels by. Metal ion leachates and the physicochemical stability of. The neutralization of glu16 negative charge by a metal ion and a decrease of the dielectric constant of the medium by addition of trifluoroethanol tfe, also promote hub aggregation. Jan 01, 2011 metalioncatalyzed oxidation depends on concentration of metal ions in the environment. Metal ions guided selfassembly of therapeutic proteins. The neutralization of glu16 negative charge by a metal ion and a decrease of the dielectric constant of the medium by addition of trifluoroethanol tfe. Physicochemical characteristics were compared, including morphology, xrd, ftir, cd, biological potency, the dissolution behaviors in vitro and plasma pharmacokinetics in vivo. Thus, it seems reasonable to speculate that ad, pd, and als share common mechanisms of metal ion regulation andor metalion induction of protein aggregation, which might lead to a common, or at least overlapping, pathogenic mechanisms. Metal ions that leach into biotherapeutic drug product solution during manufacturing and storage, result in contamination that can cause physicochemical degradation of the active molecule. Here we report that the divalent metal ion transporter dmt1, the primary nonheme iron transporter in mammals, is regulated by ubiquitination mediated by the nedd4 family member wwp2.
Aug 14, 2007 thus, it seems reasonable to speculate that ad, pd, and als share common mechanisms of metal ion regulation andor metalion induction of protein aggregation, which might lead to a common, or at least overlapping, pathogenic mechanisms. Aluminum ion, but not the others, induced neurofibrillary degeneration within the midbrain and medulla. Physicochemical characteristics were compared, including morphology, xrd, ftir, cd, biological potency, the dissolution behaviors in vitro and. Metal ion induced lateral aggregation of filamentous viruses fd and m abstract we report a detailed comparison between calculations of interfilament interactions based on montecarlo simulations and experimental features of lateral aggregation of bacteriophages fd and m induced by a number of divalent metal ions. Drugs that specifically target this process could be useful in the future therapy of these diseases.
In some metalloproteins, the coordination sphere around the metal ion is relatively static in that the coordination number and gross coordination geometry do not change as the protein executes its. Phosphorylation of the tau protein decreases the affinity of tau for metal ion binding. These results suggest that elevation of some metal ions in the brain induces protein aggregation with the nature of the aggregation being highly ion dependent. Allatom dmd features a transferable force field and has been successfully used to fold several small proteins ab initio 60 aa. Metal ioninduced lateral aggregation of filamentous viruses. Sep 14, 2012 to make a comparative study on sustained delivery performance of rhifn with random amorphous and spherical crystallike ordered selfassemblies. Metal ions participate in maturation of genomic rna, activation and catalytic mechanisms, reverse transcription, initial integration process and protection of newly synthesized dna, inhibition of proton translocation m2 protein, minus and plusstrand. Apr 16, 2014 the presence of divalent metal ions could accelerate the protein aggregation 27. There is clear evidence implicating oxidative stress in the pathology of many different neurodegenerative diseases. Salts of tungsten oxide migrated from prefilled syringes into the drug product and triggered protein oxidation followed by aggregation 33.
The generation of ros during earlystage protein aggregation may be a common, fundamental molecular mechanism underlying the pathogenesis of oxidative damage, neurodegeneration and cell death in several different neurodegenerative diseases. Determination of transition metals by ion chromatography separation. Synergistic effects of metalinduced aggregation of human serum. This is especially reflected in a decreased fibrilend elongation rate. Amyloid fibrillization is initiated by the formation of an unstable amyloidogenic intermediate which contains some structureandiscausedeitherbyipartialunfolding of a compact globular protein or ii the gain of structure by. There are several factors that should be considered in a study of mechanisms of protein stabilization. Metal ions as modulators of protein conformation and. Protein cohesion induced by metal ions observed with fluorescence correlation spectroscopy. Metal ions guided selfassembly of therapeutic proteins for.
Aggregation of biologically important peptides and proteins. A promising hypothesis is that dysfunction of metal ion homeostasis, and consequently, the undesired interaction of metal ions with a. Metal ions and redox balance regulate distinct amyloidlike. Influence of phosphorylation on tau aggregation induced by aluminium and dmso. Heavy metals and metalloids as a cause for protein. Dualaptamerconjugated molecular modulator for detecting. Furthermore, xray photoelectron spectroscopy measurement indicated that al ions might form chemical bonds with neighboring atoms and destroy the secondary structures of the protein. Protein aggregation presents a key challenge in the development of biologic formulations as it can have an impact on product quality in terms of efficacy and immunogenicity. Hydrated and weakly complexed transition metals can be separated as cations on a cation exchange column. To uncover the role of metal binding and disulfide bonding, we systematically studied the folding thermodynamics of the sod1. While most aggregates are formed during upstream operations, and the risk for aggregate formation at the cellular level has increased as titers have increased, if unsuitable conditions are selected for downstream operations, aggregation can also occur.
Metal ions are constituents of many metalloproteins, in which they have either catalytic metalloenzymes or structural functions. These interactions lead to conformational changes, which result in tau aggregation. Pdf aggregation of biologically important peptides and. In addition, we report recent advances on the role of aberrant metal coordination in promoting sod1 aggregation, highlighting the influence of metal ion homeostasis in pathologic aggregation processes.
Metal ions as modulators of protein conformation and misfolding in. Protein cohesion induced by metal ions observed with. In this study, we characterized the chemical modifications in the monoclonal antibody igg 2 aggregates generated under various conditions, including mechanical, chemical, and thermal stress treatment, to provide insight into the mechanism of protein aggregation and the types of aggregate produced by the different stresses. Dokholyan1 department of biochemistry and biophysics, university of north carolina, chapel hill, nc 27599. Biotherapeutic formulation factors affecting metal. In summary, the impact of metal ions on biologic products exhibits a complex dependence on the physicalchemical structure of the protein of interest. Jan, 2012 the highest amounts of ion leachates occurred in the formulation containing 20 mgml protein and 0. In this work, the characteristics of various metals were studied cu, zn, mg, mn, fe, co, ni, cd and ca in proteins with known crystal structure as well as the specificity of their environments. Ros reactive oxygen species are the primary mediators of oxidative stress and many of the aggregating proteins and peptides associated with neurodegenerative disease can generate hydrogen peroxide, a key ros, apparently through interactions with redoxactive metal ions. Our studies now establish that the nterminal region of a. Metal ions also bind to an additional site in the flexible region between the octarepeats and the cterminal domain. Although metal ions are too small to induce an immune response, they can conjugate with proteins to form metal protein complexes, potentially resulting in immunogenicity. For the criiiprotein conjugate antigen, albumin has been suggested to be the. Biopharmaceutical product stability considerations, part.
Recently it was shown that copper, differently from zinc, nickel. Protein aggregation, metals and oxidative stress in. Metal ioninduced lateral aggregation of filamentous. The outcomes of this research have important implications for the prediction of physiological parameters that favor aggregate formation.
Nov 15, 2008 here we report that the divalent metal ion transporter dmt1, the primary nonheme iron transporter in mammals, is regulated by ubiquitination mediated by the nedd4 family member wwp2. Dmt1 interacts with 2 ww domaininteracting proteins, ndfip1 and ndfip2, previously proposed to have roles in protein trafficking. Our mission is to help scientists accelerate discovery by operating a platform for research communication that encourages and recognises the most responsible behaviours in science. Our previous studies established a hierarchy in as. Dynamical roles of metal ions and the disulfide bond in cu. Determination of transition metals by ion chromatography. Challenges of protein aggregation during purification. Molecules free fulltext the role of metal binding in the. To make a comparative study on sustained delivery performance of rhifn with random amorphous and spherical crystallike ordered selfassemblies. Although many players are involved in the in vivo protein aggregation, physiological factors such as labile metal ions within the cellular environment are likely to play a key role. Pdf the study of the aggregation processes in presence of metal ions is an essential step for understanding the key role of metals in proteinprotein. Metal ioninduced lateral aggregation of filamentous viruses fd and m jay x.
Prosthetic groups small organic molecule or metal ion associated with a protein o regions of secondary structure interact to give a protein it tertiary structure. The role of metals in protein conformational disorders the case of. Aggregation of biologically important peptides and. Chemical modifications in therapeutic protein aggregates.
Pdf thermal aggregation of proteins in the presence of. In some metalloproteins, the coordination sphere around the metal ion is relatively static in that the coordination number and gross coordination geometry do not change as the protein executes its function. Biological processes are often metal ion specific, although more than one metal ion can play each of these roles. To study the effect of metal ions on the aggregation of hiapp peptides, alcl 3, and zncl 2 were added respectively into peptide solution to a desired ion concentration e. Developing biosensing probes for bioactive metal ion detection can contribute to fields including disease diagnosis and therapy and studying the mechanisms of biological activities. Protein aggregation and neurodegenerative diseases. The mechanism of protein stabilization in the solid state is still a controversial topic. Pdf thermal aggregation of proteins in the presence of metal ions. Mechanistic basis of the inhibition of slc11nrampmediated metal ion transport by bisisothiourea substituted compounds. Neurodegenerative disorders share common features comprising aggregation of misfolded proteins, failure of the ubiquitinproteasome system, and increased levels of metal ions in the brain. Highlights metal ions play a decisive role in the regulation of protein function and folding but can also promote protein misfolding and aggregation. It can also be nonbiospecific, for example, a protein binding dye substance or histidinecontaining proteins binding metal ions as in immobilized metal ion affinity chromatography, which will be described in a separate section due to its importance. Some negatively charged carboxylic acid groups also attract positively charged amine groups through electrostatic. Here we have shown that zinc and copper ions differentially affect.
Furthering our understanding of the relationship between metal ion concentration and protein aggregation will prove useful for future scientific applications, such as finding therapies for. In order to evaluate the influence of protein phosphorylation on the effects of these aggregation inducers, we employed gsk3. By binding several protein side chains, metals can act as multidentate crosslinking agents. Synergistic influence of phosphorylation and metal ions on. Henry membrane transport group, department of chemistry, the faculties, australian national university, canberra city, act, australia. The role of metal binding in the amyotrophic lateral. Iron has been implicated in the aggregation of proteins responsible for the pathophysiology of several neurodegenerative diseases, such as alzheimer and parkinsons 15. Transition metal ions, like cu, zn and fe are implicated in many neurodegenerative disorders and are believed to influence either the aggregation of amyloidogenic proteins or the formation of reactive oxygen species ros. Under normal conditions, a is rarely produced in the brain, since amyloid precursor protein app is cleaved by and secretase. Thermal aggregation of lactoglobulin in presence of metal. Though aging is considered as a prominent risk factor for ad pathogenesis, substantial evidence suggests that an imbalance of essential biometal ions in the body and exposure to certain metal ions in the environment can. Oct 21, 2019 either demetallation or aberrant metal binding has been shown to promote protein misfolding and pathogenic aggregation 23,38,39. The metal ion bound state appears to be incapable of aggregation, effectively reducing the available monomeric a.
Formulation in sucrose can increase aggregation over time because of protein glycation when sucrose is hydrolyzed. The impact of common formulation factors such as protein concentration, antioxidant, metal chelator concentration and type, surfactant, ph, and contact time with stainless steel on metal leachables was investigated by a design of experiments. Introduction the rate of aggregation of proteins depends strongly on the concentration of the aggregating proteins, but this relationship. Role of metal ions in aggregation of intrinsically disordered proteins. The presence of divalent metal ions could accelerate the protein aggregation 27.
The presence of certain ligands including certainions may enhance aggregation. The study of the aggregation processes in presence of metal ions is an essential step for understanding the key role of metals in proteinprotein and proteinsolvent interactions. Regulation of the divalent metal ion transporter dmt1 and. Under pathological conditions, app is cleaved with and secretase, generating neurotoxic a peptide. West indian medical journal university of the west indies.
Bioactive metal ions play important roles in both physiological and pathological processes. In this framework, the presence of metal ions in the protein solutions can lead to different aggregation andor denaturation processes respect to the one. Protein aggregates within affected cells often contain ubiquitin, however no report has focused on the aggregation propensity of this protein. Journal of environmental science and health, part a. Removal of protein aggregates from biologic apis is crucial due to their potential to increase immunogenicity. Metal ions play a decisive role in the regulation of protein function and folding but can also promote protein misfolding and aggregation. This study evaluated the impact of elevated aluminum, ferric, calcium and magnesium ions on rabbit brain. Thermal aggregation of lactoglobulin in presence of metal ions. Engineering metal ion coordination to regulate amyloid. Our studies could deepen the understanding of the role of metal ions in the aggregation of amyloid peptides.
Zinc, magnesium and copper are the commonest metal ion that binds with viral proteins. Protein aggregation, ion channels, and membrane damage croat med j 2001. Dynamical roles of metal ions and the disulfide bond in cu, zn superoxide dismutase folding and aggregation feng ding and nikolay v. However, at least some of the difficulties dealing with this topic are due to inadequate tools andor incomplete analysis of the data. Interactions with metal surfaces can lead to epitaxic denaturation, which triggers aggregate formation. By interfering with the folding process, heavy metal ions and. Laguna designgetty images biopharmaceutical manufacturers are under increasing pressure from regulators to ensure the safety and quality of their products. Biotherapeutic formulation factors affecting metal leachables.
While most aggregates are formed during upstream operations, and the risk for aggregate formation at the cellular level has increased as titers have increased, if unsuitable conditions are selected for downstream operations, aggregation can also occur during downstream processing. Mechanisms of protein stabilization in the solid state. View enhanced pdf access article on wiley online library. Tau protein aggregation and its hyperphosphorylation play an important role in the pathogenesis of alzheimers disease. The neutralization of glu16 negative charge by a metal ion and a decrease of the dielectric constant of the medium by addition of. Increasing mechanical strength of gelatin hydrogels by divalent metal ion removal. It is sufficient to shift the same mutation by only two residues e18v to regain the behavior of wild. There are limited reports on the effect of or understanding of the mechanism of stabilization by amino acids in solid state. Amyloid and tau aggregation are implicated in manifold neurodegenerative diseases and serve as two signature pathological hallmarks in alzheimers disease ad. Molecules free fulltext the role of metal binding in. Protein purification methods process development forum. It was found that all metal ions induced hsa aggregation, and this effect was.
Engineering metal ion coordination to regulate amyloid fibril. Trace amounts of metals are inevitably present in biotherapeutic products. Implications for ion channel formation and membrane damage joseph i. Metal ion, aggregation, oxidation, protein, biotherapeutics. Global 3dimensional arrangement of all atoms in a protein o includes. On the other hand, the other three ions caused aggregation of low molecular weight mitochondrial proteins. Protein misfolding and conformational changes are common hallmarks in many neurodegenerative diseases involving formation and deposition of toxic protein aggregates. Analyzing protein aggregation in biopharmaceuticals. The rhifn selfassemblies were identified in batch crystallization mode. Though aging is considered as a prominent risk factor for ad pathogenesis, substantial evidence suggests that an imbalance of essential biometal ions in the body and exposure to certain metal ions in the environment can potentially. High levels of labile metal ions in the synaptic milieu are likely to facilitate protein aggregation in. Furthering our understanding of the relationship between metal ion concentration and protein aggregation will prove useful for future scientific. What are the causes of protein aggregation in biologic formulations. Neurodegenerative disorders are the pathological conditions in which cells of the brain andor spinal cord.
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