Cell Adh Migr. 2011 Mar 1;5(2):5-7. [Epub ahead of print]
Extracellular matrix-derived peptides and myocardial repair.
Cardiovascular Research Institute; University of California San Francisco; San Francisco, CA USA.
Abstract
Repairing cardiac tissue remains one of the most challenging goals in tissue engineering. Here, we discuss ways whereby we sought to treat myocardial infarctions using extracellular-matrix derived peptides. Using an ischemia/reperfusion myocardial infarction rodent model, we targeted these extracellular matrix-derived peptides to the myocardial infarct site and were able to induce angiogenesis and alter the negative remodeling seen after an acute myocardial infarction. Our results indicate a potentially new strategy for repairing damaged tissue.
PMID: 21048428 [PubMed - as supplied by publisher]
J Pharm Biomed Anal. 2011 Feb 20;54(3):572-6. Epub 2010 Sep 16.
Evaluation of a synthetic peptide as a replacement for the recombinant fusion protein of respiratory syncytial virus in a potency ELISA.
McGivney JB 4th, Bishop E, Miller K, Casas-Finet J, Yang H, Wei Z, Strouse R, Schenerman M.
Department of Analytical Biochemistry, MedImmune, One MedImmune Way, Gaithersburg, MD 20878, United States.
Abstract
This report describes the development of a potency ELISA using a peptide derived from the motavizumab binding epitope of respiratory syncytial virus (RSV) F-protein. Motavizumab is an antibody therapeutic studied for the prevention of RSV disease. It binds to the RSV glycoprotein F (F-protein), blocking the ability of RSV to fuse with target cells. This binding is the basis for a potency ELISA, however, due to inefficient F-protein production, development of an alternative ligand for the potency ELISA was investigated. A series of synthetic peptides spanning the motavizumab epitope on F-protein were evaluated for motavizumab binding activity. A 26-mer peptide was identified with desirable motavizumab binding kinetics, as shown by ELISA and surface plasmon resonance. The peptide corresponds to a portion of the motavizumab binding domain on the F-protein, and is referred to as F-peptide. The binding of motavizumab to the F-peptide is used in a new motavizumab potency ELISA, which was shown to be robust and statistically comparable to the F-protein ELISA. In addition, based on a qualitative observation, this new ELISA may be able to detect motavizumab degradation with greater sensitivity compared to the F-protein ELISA.
Copyright © 2010 Elsevier B.V. All rights reserved.PMID: 20943340 [PubMed - in process]
Clin Sci (Lond). 2011 Feb;120(3):99-120.
Vitiligo, reactive oxygen species and T-cells.
Division of Dermatology, Department of Medicine, University of Ottawa, Ottawa, Ontario, Canada. sglassman@toh.on.ca
Abstract
The acquired depigmenting disorder of vitiligo affects an estimated 1% of the world population and constitutes one of the commonest dermatoses. Although essentially asymptomatic, the psychosocial impact of vitiligo can be severe. The cause of vitiligo remains enigmatic, hampering efforts at successful therapy. The underlying pathogenesis of the pigment loss has, however, been clarified to some extent in recent years, offering the prospect of effective treatment, accurate prognosis and rational preventative strategies. Vitiligo occurs when functioning melanocytes disappear from the epidermis. A single dominant pathway is unlikely to account for all cases of melanocyte loss in vitiligo; rather, it is the result of complex interactions of biochemical, environmental and immunological events, in a permissive genetic milieu. ROS (reactive oxygen species) and H2O2 in excess can damage biological processes, and this situation has been documented in active vitiligo skin. Tyrosinase activity is impaired by excess H2O2 through oxidation of methionine residues in this key melanogenic enzyme. Mechanisms for repairing this oxidant damage are also damaged by H2O2, compounding the effect. Numerous proteins and peptides, in addition to tyrosinase, are similarly affected. It is possible that oxidant stress is the principal cause of vitiligo. However, there is also ample evidence of immunological phenomena in vitiligo, particularly in established chronic and progressive disease. Both innate and adaptive arms of the immune system are involved, with a dominant role for T-cells. Sensitized CD8+ T-cells are targeted to melanocyte differentiation antigens and destroy melanocytes either as the primary event in vitiligo or as a secondary promotive consequence. There is speculation on the interplay, if any, between ROS and the immune system in the pathogenesis of vitiligo. The present review focuses on the scientific evidence linking alterations in ROS and/or T-cells to vitiligo.
PMID: 20958268 [PubMed - in process]
Epigenetics. 2011 Feb 1;6(2). [Epub ahead of print]
Detailed specificity analysis of antibodies binding to modified histone tails with peptide arrays.
Bock I, Dhayalan A, Kudithipudi S, Brandt O, Rathert P, Jeltsch A.
Biochemistry Laboratory, School of Engineering and Science, Jacobs University Bremen, Bremen, Germany.
Abstract
Chromatin structure is greatly influenced by histone tail post-translational modifications (PTM), which also play a central role in epigenetic processes. Antibodies against modified histone tails are central research reagents in chromatin biology and molecular epigenetics. We applied Celluspots peptide arrays for the specificity analysis of 36 commercial antibodies from different suppliers which are directed towards modified histone tails. The arrays contained 384 peptides from eight different regions of the N-terminal tails of histones, viz. H3 1-19, 7-26, 16-35 and 26-45, H4 1-19 and 11-30, H2A 1-19 and H2B 1-19, featuring 59 post-translational modifications in many different combinations. Using various controls we document the reliability of the method. Our analysis revealed previously undocumented details in the specificity profiles of the tested antibodies. Most of the antibodies bound well to the PTM they have been raised for, but some failed. In addition, some antibodies showed high cross-reactivity and most antibodies were inhibited by specific additional PTMs close to the primary one. Furthermore, specificity profiles for antibodies directed toward the same modification sometimes were very different. The specificity of antibodies used in epigenetic research is an important issue. We provide a catalog of antibody specificity profiles for 36 widely used commercial histone tail PTM antibodies. Better knowledge about the specificity profiles of antibodies will enable researchers to implement necessary control experiments in biological studies and allow more reliable interpretation of biological experiments using these antibodies.
PMID: 20962581 [PubMed - as supplied by publisher]
Cancer Biol Ther. 2011 Jan 29;11(1). [Epub ahead of print]
Identification of a high affinityTAG-72 binding peptide by phage display selection.
Xiao N, Cheng D, Wang Y, Chen L, Liu X, Dou S, Liu G, Liang M, Hnatowich DJ, Rusckowski M.
Department of Radiology, University of Massachusetts Medical School, Worcester, Massachusetts, USA.
Abstract
Purpose: Phage display was used to select novel peptides that specifically bind the TAG-72 antigen and with properties suitable for imaging TAG-72 positive cancers. Results: After three rounds of selection against TAG-72 and using two different elution conditions including a long elution, the consensus sequences FRE RCD KHP QKC TKF L and DPR HCQ KRV LPC PAW L were expressed on phages G3-15 and T3-15 respectively. ELISA , fluorescence-activated cell sorting analysis and fluorescence microscopy provided evidence that both phages specifically bound TAG-72 in vitro. Both peptides are stable in 37°C serum. By a cell binding competition assay, the IC(50) for T3-15 was measured as 0.29 nM and therefore 36-fold higher affinity than G3-15 at 10.32 nM. The biodistribution in mice carrying LS-174T tumors in one thigh were similar for both (99m)Tc-peptides at 30 min, but at 90 min the (99m)Tc-T3-15 peptide accumulated almost three times higher in the tumor. The SPE CT/CT images were consistent with the biodistribution results. Procedures: The f88-4/Cys6 phage library and two different elution conditions were used to identify two new higher affinity binding peptides for the TAG-72 antigen. One, was a single brief elution with pH 2.2 glycine buffer and the second began with the glycine elution but was followed with a longer elution with Tris buffered saline (TBS) at pH 7.4. The phages that bound TAG-72 were evaluated by fluorescence-activated cell sorting analysis using TAG-72 positive LS-174T cells and confirmed by immunofluorescence imaging. The consensus peptides displayed on the selected phages were synthesized and conjugated with NHS -MAG3 for radiolabeling with (99m)Tc. The IC(50) for TAG-72 binding was evaluated by cell binding competition in vitro while binding affinity was evaluated in vivo by necropsy and SPE CT/CT imaging in a tumor mouse model. Conclusion: We have identified a peptide with a sub nanomolar inhibition constant for the TAG-72 antigen that may have applications in cancer imaging.
PMID: 20980835 [PubMed - as supplied by publisher]
J Pharm Biomed Anal. 2011 Jan 25;54(2):303-10. Epub 2010 Sep 24.
Stability of Myrmecia pilosula (Jack Jumper) Ant venom for use in immunotherapy.
Wiese MD, Davies NW, Chataway TK, Milne RW, Brown SG, Heddle RJ.
School of Pharmacy and Medical Sciences, University of South Australia, GPO Box 2471, Adelaide, South Australia 5001, Australia. Michael.wiese@unisa.edu.au
Abstract
Allergy to Myrmecia pilosula (Jack Jumper Ant) venom is common in Australia, affecting ∼2.7% of some communities. Venom immunotherapy is a highly effective treatment, but for the venom to be widely distributed for clinical use, the stability and shelf-life of formulated Jack Jumper Ant venom must be demonstrated. HPLC-UV, ELISA Inhibition, SDS-PAGE and SDS-PAGE Immunoblot were used to assess venom stability under conditions of varying temperature, pH and in the presence of various stabilising agents. Optimal stability occurred between pH 8 and 10, however the presence of benzyl alcohol within this pH range resulted in a cloudy appearance within 3 days, so a pH of 6 was used. Increasing polysorbate 80 concentrations accelerated the degradation of allergenic peptides in 100 μg/mL venom, but improved stability at concentrations of 1 μg/mL or less. Sucrose reduced degradation of allergens Myr p 1 and Myr p 3, whilst glycerol was destabilizing. In the presence of 22% sucrose, 1.1mg/mL Jack Jumper Ant venom was stable at -18 °C and 4 °C for 12 months; following dilution to 100 μg/mL with 0.9% sodium chloride, 10mM phosphate (pH 6), 0.05% polysorbate 80 and 0.9% benzyl alcohol (giving 2% sucrose), venom was stable for 7 days when stored at 4 °C. Concentrated Jack Jumper Ant venom can be stored in 22% sucrose for 12 months, and after dilution to 100 μg/mL for clinical use, it should be discarded after 7 days.
Copyright © 2010 Elsevier B.V. All rights reserved.PMID: 20869831 [PubMed - in process]
Behav Brain Res. 2011 Jan 20;216(2):712-8. Epub 2010 Sep 29.
Dipeptidyl peptidase IV (DPP4)-deficiency attenuates diet-induced obesity in rats: Possible implications for the hypothalamic neuropeptidergic system.
Stephan M, Radicke A, Leutloff S, Schmiedl A, Pabst R, von Hörsten S, Dettmer S, Lotz J, Nave H.
Clinic for Psychosomatics and Psychotherapy, Hannover Medical School, OE 7160, Carl-Neuberg-Strasse 1, 30625 Hannover, Germany.
Abstract
The underlying mechanisms controlling food intake and satiety are thoroughly controlled, but seem to be insufficient under conditions of almost unlimited food supply. Hence, overweight and obesity are serious problems especially in industrialized countries. To assess the possible influence of CD26, exerting a dipeptidyl peptidase activity (DPP4) cleaving several energy homeostasis-relevant peptides, we investigated wild type and DPP4-deficient dark agouti rats in a model of diet-induced obesity and found a reduced weight gain in DPP4-deficient rats. When investigating the specific increase of whole body fat volume by MRI to assess the distribution pattern (subcutaneous vs. intraabdominal), there was an altered ratio under dietary conditions only in DPP4-deficient rats, which was due to lower intraabdominal fat amounts. Furthermore, we investigated the number of cells immunopositive for the leptin receptor (OB-R), the orexigenic leptin antagonist neuropeptide Y (NPY), as well as of the NPY receptors Y1, Y2, and Y5 within hypothalamic nuclei. Independent from the body weight, higher levels of NPY and all receptors were expressed in DPP4-deficent rats. Under obese conditions, hypothalamic Y2-levels were reduced in both strains. Concerning NPY and Y1, there were partly oppositional effects, with reduced hypothalamic Y1 levels only in wild types, and increased NPY levels only in DPP4-deficient rats. These effects might be responsible for unaltered food intake in DPP4-deficent rats compared to wild types, despite reduced weight gain. However, since the food intake remained unaffected, these effects suggest that DPP4 exerts its effects on intraabdominal fat also via peripheral actions.
Copyright © 2010 Elsevier B.V. All rights reserved.PMID: 20887754 [PubMed - in process]
Anal Biochem. 2011 Jan 1;408(1):37-45. Epub 2010 Sep 20.
Enhancing the stability of ¹⁸O-labeled peptides through removal of immobilized trypsin by ZipTips.
Li MY, Peng F, Zuo JH, Yi H, Tang CE, Li C, Zhang PF, Chen ZC, Xiao ZQ.
Xiangya Hospital, Central South University, Changsha, China.
Abstract
Trypsin-catalyzed ¹⁸O labeling is increasingly used in shotgun proteomics for relative peptide/protein quantitation. However, precise quantitative measurements are often complicated by the instability of ¹⁸O-labeled peptides caused mainly by oxygen back-exchange. Although a number of attempts have been made to reduce or prevent oxygen back-exchange, there is still room for improvement. Here we demonstrate that the removal of immobilized trypsin by filtration using ZipTips can efficiently minimize oxygen back-exchange and enhance the stability of ¹⁸O-labeled peptides under various pH conditions. The ¹⁸O-labeled peptides processed by the approach were successfully separated by immobilized pH gradient-isoelectric focusing (IPG-IEF), and no marked decrease in the extent of labeling was observed. The results also demonstrated that there was no correlation between the extent of ¹⁸O labeling and molecular weight or isoelectric point (pI). The approach presented here is especially applicable to microscale samples. Its ability to generate stably ¹⁸O-labeled samples without back-exchange should expand the application scope of the ¹⁸O-labeling technique.
Copyright © 2010 Elsevier Inc. All rights reserved.PMID: 20816659 [PubMed - in process]
Anal Biochem. 2011 Jan 1;408(1):105-17. Epub 2010 Aug 31.
Mass spectrometric characterization of the isoforms in Escherichia coli recombinant DNA-derived interferon alpha-2b.
Liu YH, Wylie D, Zhao J, Cure R, Cutler C, Cannon-Carlson S, Yang X, Nagabhushan TL, Pramanik BN.
Merck Research Laboratories, Kenilworth, NJ 07033, USA. yan-hui.liu@merck.com
Abstract
The isoforms Iso-2, Iso-3, and Iso-4 of Escherichia coli-derived recombinant human interferon alpha-2b (rhIFN α-2b), generated by posttranslational modifications of the protein during fermentation, present a major problem in terms of purification and the yield of the drug substance. We report here the structural characterization of these isoforms by mass spectrometry (MS) methods. An extensive MS study was conducted on Iso-4, which is composed of up to 75% of the in-process IFN, and on the native rhIFN α-2b. The trypsin-digested peptide mixtures generated from the two samples were analyzed by liquid chromatography (LC)-MS, and targeted peptides were further studied by LC-tandem MS (triple quadrupole mass spectrometer), high-resolution MS(n) (LTQ Orbitrap), and matrix-assisted laser desorption/ionization MS (MALDI-MS). The structure of Iso-4 was elucidated as a novel pyruvic acid ketimine derivative of the N-terminal cysteine (Cys1) of IFN α-2b, where the disulfide bond between Cys1 and Cys98 was fully reduced and the other disulfide bond pair, Cys29-ss-Cys138, was partially reduced. Similarly, Iso-2 was identified as a correctly disulfide-folded rhIFN α-2b with acetylation on Cys1, and Iso-3 was identified as an S-glutathionylated form (Cys98) of partially reduced rhIFN α-2b that was pyruvated on Cys1. Based on the characterization work, a reproducible conversion procedure was successfully implemented to convert Iso-4 to rhIFN α-2b.
Copyright © 2010 Elsevier Inc. All rights reserved.PMID: 20807495 [PubMed - in process]
Anal Biochem. 2011 Jan 1;408(1):136-46. Epub 2010 Sep 17.
Label-free liquid chromatography-tandem mass spectrometry analysis with automated phosphopeptide enrichment reveals dynamic human milk protein phosphorylation during lactation.
Froehlich JW, Chu CS, Tang N, Waddell K, Grimm R, Lebrilla CB.
Department of Chemistry, University of California, Davis, 95616, USA.
Abstract
Protein phosphorylation is a critical posttranslational modification that affects cell-cell signaling and protein function. However, quantifying the relative site-specific changes of phosphorylation occupancies remains a major issue. An online enrichment of phosphopeptides using titanium dioxide incorporated in a microchip liquid chromatography device was used to analyze trypsin-digested human milk proteins with mass spectrometry. The method was validated with standards and used to determine the dynamic behavior of protein phosphorylation in human milk from the first month of lactation. α-Casein, β-casein, osteopontin, and chordin-like protein 2 phosphoproteins were shown to vary during this lactation time in an independent manner. In addition, changes in specific regions of these phosphoproteins were found to vary independently. Novel phosphorylation sites were discovered for chordin-like protein 2, α-lactalbumin, β-1,4-galactosyl transferase, and poly-Ig (immunoglobulin) receptor. Coefficients of variation for the quantitation were comparable to those in other contemporary approaches using isotopically labeled peptides, with a median value of 11% for all phosphopeptide occupancies quantified.
Copyright © 2010 Elsevier Inc. All rights reserved.PMID: 20804719 [PubMed - in process]PMCID: PMC2964395 [Available on 2012/1/1]
Anal Biochem. 2011 Jan 1;408(1):19-31. Epub 2010 Aug 7.
An optimized magnetite microparticle-based phosphopeptide enrichment strategy for identifying multiple phosphorylation sites in an immunoprecipitated protein.
Huang Y, Shi Q, Tsung CK, Gunawardena HP, Xie L, Yu Y, Liang H, Yang P, Stucky GD, Chen X.
Department of Chemistry, Institutes of Biomedical Sciences, Fudan University, Shanghai, People's Republic of China.
Abstract
To further improve the selectivity and throughput of phosphopeptide analysis for the samples from real-time cell lysates, here we demonstrate a highly efficient method for phosphopeptide enrichment via newly synthesized magnetite microparticles and the concurrent mass spectrometric analysis. The magnetite microparticles show excellent magnetic responsivity and redispersibility for a quick enrichment of those phosphopeptides in solution. The selectivity and sensitivity of magnetite microparticles in phosphopeptide enrichment are first evaluated by a known mixture containing both phosphorylated and nonphosphorylated proteins. Compared with the titanium dioxide-coated magnetic beads commercially available, our magnetite microparticles show a better specificity toward phosphopeptides. The selectively-enriched phosphopeptides from tryptic digests of β-casein can be detected down to 0.4 fmol μl⁻¹, whereas the recovery efficiency is approximately 90% for monophosphopeptides. This magnetite microparticle-based affinity technology with optimized enrichment conditions is then immediately applied to identify all possible phosphorylation sites on a signal protein isolated in real time from a stress-stimulated mammalian cell culture. A large fraction of peptides eluted from the magnetic particle enrichment step were identified and characterized as either single- or multiphosphorylated species by tandem mass spectrometry. With their high efficiency and utility for phosphopeptide enrichment, the magnetite microparticles hold great potential in the phosphoproteomic studies on real-time samples from cell lysates.
Published by Elsevier Inc.PMID: 20696126 [PubMed - in process]
Appl Radiat Isot. 2011 Jan;69(1):68-74. Epub 2010 Sep 15.
Radiopeptide internalisation and externalization assays: cell viability and radioligand integrity.
Naqvi SA, Sosabowski JK, Nagra SA, Ishfaq MM, Mather SJ, Matzow T.
Centre for Molecular Oncology and Imaging, Institute of Cancer, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, Charterhouse Square, London EC1M 6BQ, United Kingdom.
Abstract
Various aspects of radiopeptide receptor-mediated cell internalisation and externalization assays were assessed, including the integrity of externalized peptides and the effect of varying the pH and incubation time of the acid wash step (to remove surface receptor-bound ligand) on efficacy and cell viability. The observed intact proportion of externalized peptide was 5-10%, and acid wash buffers with pH 2.8 or below were found to be detrimental to cell viability and integrity, particularly following prolonged incubation times.
Copyright © 2010 Elsevier Ltd. All rights reserved.PMID: 20880713 [PubMed - in process]
Appl Radiat Isot. 2011 Jan;69(1):52-5. Epub 2010 Sep 8.
Enhancement of reaction conditions for the radiolabelling of DOTA-peptides with high activities of yttrium-90.
Nardelli A, Castaldi E, Ortosecco G, Speranza A, Storto G, Pace L, Salvatore M.
Istituto di Biostrutture e Bioimmagini, Consiglio Nazionale delle Ricerche (CNR), Naples, Italy. a.nardelli@libero.it
Abstract
Peptide receptor radionuclide therapy (PRRT) has recently expanded due to radiolabelling of DOTA-peptides, such as the somatostatin analogues [DOTA(0), Tyr(3)]octreotate (DOTATATE). The achievement of high specific activities during procedures has been indicated as the critical factor to consent effective therapy. Several radiochemical factors may negatively impact reaction procedures such as pH, temperature and time of reaction. Our study was undertaken to explore the influence of radiochemical parameters, such as time of incubation, on reaction kinetics during the radiolabelling of DOTATATE with (90)Y.
METHODS: Forty-five radiolabelling procedures were carried out using small volumes of yttrium-90, typically 60-78 μL. At nearly constant pH and temperature two different settings of radiolabelling procedures were implemented, removing the products from the heating water bath approximately after 30 min (group E, early; n=20) and after 39 min (group L, later; n=25). Quality controls were performed by means of both high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC).
RESULTS: Reaction kinetics for (90)Y were found to a provide suitable percentage of incorporation at pH 4.5 for both groups. Reaction temperature was not different between groups E and L. A significant difference was found between the two groups in radiochemical yield, which was 95.6% ± 0.8 for group E and 98.2% ± 1.1 for group L (p<0.0001).>
CONCLUSION: In order to achieve optimal specific activities, pH, temperature and time of reaction necessitate careful evaluation and setting. A statistically significant difference in labelling yield was found between a set of procedures completed at 39 min as compared to that executed at 30 min, keep the reaction pH and temperature constant.
Copyright © 2010 Elsevier Ltd. All rights reserved.PMID: 20869877 [PubMed - in process]
Biomaterials. 2011 Jan;32(1):249-257. Epub 2010 Sep 28.
A peptide-based material platform for displaying antibodies to engage T cells.
Zheng Y, Wen Y, George AM, Steinbach AM, Phillips BE, Giannoukakis N, Gawalt ES, Meng WS.
Division of Pharmaceutical Sciences, Duquesne University, Pittsburgh, PA 15282, USA.
Abstract
This study investigated a strategy by which antibodies are displayed to engage T cells. A peptide composite containing binding sites was characterized in vitro and explored as an injectable system in vivo. The composite consists of two amphiphilic peptides, AEAEAKAKAEAEAKAK (referred to as "EAK") and EAK appended with six consecutive histidines at the C-terminus ("EAKH6"). Spectroscopic analysis showed the two peptides integrated into a single structure. Prior to combination, conformational analysis revealed that EAKH6 adopts a mixed α-helix/β-strand conformation. In the presence of EAK, EAKH6 exists predominately in β-strand conformation. The composite of EAK-EAKH6 was found to display His-tags, using nickel-bound horseradish peroxidase as a probe. T cell-specific antibodies were found stably displayed on the EAK-EAKH6 assembly using recombinant protein A/G and anti-histidines antibody as an adaptor. When mounted with anti-CD4 antibody, the system was shown to capture CD4T cells in a mixed population of lymphocytes. Antibodies were concentrated in the subcutaneous space in mice when co-administered with EAK and EAKH6 along with protein A/G and anti-histidines antibody as a solution. We report here the use of amphiphilic peptides to display Ab in vivo, the results indicating that the design can be used as a platform for engaging specific subsets of leukocytes for the purpose of immune modulation.
Copyright © 2010 Elsevier Ltd. All rights reserved.PMID: 20880580 [PubMed - as supplied by publisher]
Biomaterials. 2011 Jan;32(1):231-238.
Post-formulation peptide drug loading of nanostructures for metered control of NF-κB signaling.
Pan H, Ivashyna O, Sinha B, Lanza GM, Ratner L, Schlesinger PH, Wickline SA.
Department of Medicine, Washington University School of Medicine, St Louis, MO, USA.
Abstract
The NF-κB signaling pathway is an attractive therapeutic target for cancer and chronic inflammatory diseases. In this study, we report the first strategy to achieve NF-κB inhibition with a peptide inhibitor loaded into perfluorocarbon nanoparticles with the use of a simple post-formulation mixing approach that utilizes an amphipathic cationic fusion peptide linker strategy for cargo insertion. A stable peptide-nanoparticle complex is formed (dissociation constant ∼0.14 μM) and metered inhibition of both NF-κB signaling and downstream gene expression (ICAM-1) is demonstrated in leukemia/lymphoma cells. This post-formulation cargo loading strategy enables the use of a generic synthetic or biologic lipidic nanostructure for drug conjugation that permits flexible specification of types and doses of peptides and/or other materials as diagnostic or therapeutic agents for metered incorporation and cellular delivery.
Copyright © 2010 Elsevier Ltd. All rights reserved.PMID: 20864161 [PubMed - as supplied by publisher]
Biomaterials. 2011 Jan;32(1):222-230. Epub 2010 Sep 22.
A protein transduction domain located at the NH(2)-terminus of human translationally controlled tumor protein for delivery of active molecules to cells.
Kim M, Kim M, Kim HY, Kim S, Jung J, Maeng J, Chang J, Lee K.
College of Pharmacy, Center for Cell Signaling & Drug Discovery Research, Ewha Womans University, Seoul, Republic of Korea.
Abstract
Protein transduction domains (PTDs) are small peptides, able to penetrate biological membranes and deliver various types of cargo both in vitro and in vivo. Because use of PTDs originating from viral origins resulted in undesired effects, PTDs originating from non-viral origins are needed. Here, we report that a 10-amino acid peptide (MIIYRDLISH) derived from the NH(2)-terminus of human translationally controlled tumor protein (TCTP) functions as a PTD. This peptide was internalized through lipid raft-dependent endocytosis and partial macropinocytosis, and did not enter lysosome and nucleus. Beta-galactosidase fused to TCTP-PTD, when injected into mice, was efficiently delivered to liver, kidney, spleen, heart, and lungs of the animals. Preincubation of TCTP-PTD with adenovirus increased adenoviral mediated-gene expression in cells and also improved immune response to intranasally administered adenovirus expressing the triple repeat of G glycoprotein of respiratory syncytial virus (RSV), rAd/3×G. These findings suggest that TCTP-PTD might overcome the limitations of polycation-mediated transduction and serve as an efficient vehicle for drug delivery.
Copyright © 2010 Elsevier Ltd. All rights reserved.PMID: 20863558 [PubMed - as supplied by publisher]
Biopolymers. 2011 Jan;95(1):62-9.
Interruption of a 3(10)-helix by a single Gly residue in a poly-Aib motif: A crystallographic study.
Solà J, Helliwell M, Clayden J.
School of Chemistry, University of Manchester, Manchester M13 9PL, UK.
Abstract
The structural influence of a single Gly residue inserted into an Aib(16) homooligomer was studied in the solid state by X-ray crystallography. The peptides N(3)Aib(8)GlyAib(8)PheNH(2) (1) and CbzPheAib(8)GlyAib(8) (2) were found to adopt well-defined helical structures, which are broadly 3(10) helical. Indeed, 2 is the longest crystallographic 3(10) helix thus far reported. However, in the region of the central Gly residue, a loosening of the 3(10) structure is observed in both peptides, with 1 clearly showing local adoption of an α-helical structure in the region of residues 7-9. © 2010 Wiley Periodicals, Inc. Biopolymers 95: 62-69, 2011.
PMID: 20725951 [PubMed - in process]
Clin Dev Immunol. 2011;2011:681956. Epub 2010 Nov 1.
Behçet's Disease (Adamantiades-Behçet's Disease).
Kaneko F, Togashi A, Saito S, Sakuma H, Oyama N, Nakamura K, Yokota K, Oguma K.
Institute of Dermato-Immunology and Allergy, Southern TOHOKU Research Institute for Neuroscience, 7-115 Yatsuyamada, Koriyama, Fukushima 963-8563, Japan.
Abstract
Adamantiades-Behçet's disease (ABD) is characterized by starting with oral aphthous ulceration and developing of the systemic involvements. The pathogenesis of ABD is closely correlated with the genetic factors and the triggering factors which acquire delayed-type hypersensitivity reaction against oral streptococci mediated by IL-12 cytokine family. HLA-B51 is associated in more than 60% of the patients and its restricted CD8+ T cell response is clearly correlated with the target tissues. Bes-1 gene encoded partial S. sanguinis genome which is highly homologous with retinal protein, and 65 kD heat shock protein (Hsp-65) released from streptococci is playing an important role with human Hsp-60 in the pathogenesis of ABD. Although Hsp-65/60 has homologies with the respective T cell epitope, it stimulates peripheral blood mononuclear cells (PBMCs) from ABD patients. On the other hand, some peptides of Hsp-65 were found to reduce IL-8 and IL-12 production from PBMCs of ABD patients in active stage.
PMID: 21052488 [PubMed - in process]PMCID: PMC2967828Free PMC Article
Dev Comp Immunol. 2011 Jan;35(1):44-50. Epub 2010 Aug 13.
The effect of nitric oxide and hydrogen peroxide in the activation of the systemic immune response of Anopheles albimanus infected with Plasmodium berghei.
Herrera-Ortiz A, Martínez-Barnetche J, Smit N, Rodriguez MH, Lanz-Mendoza H.
Centro de Investigaciones sobre enfermedades Infecciosas, I.N.S.P., Avenida Universidad No. 655, Col. Santa María Ahuacatitlán, Cuernavaca, Morelos C.P. 62100, Mexico.
Abstract
The expression of genes encoding the antimicrobial peptides (AMPs) attacin, cecropin and gambicin, as well as the effects of NO and H(2)O(2) on their expression was investigated in midguts and fat bodies of Anopheles albimanus during the midgut infection with Plasmodium berghei. Midgut infection induced an increase in the expression of the three AMPs in both tissues; while NO and H(2)O(2) were present in haemolymph. Treatment with L-NAME and vitamin C reduced the effect of P. berghei infection on the AMP's expression, and exogenous NO and H(2)O(2) induced their expression in the mosquito fat body. The induction of AMPs in abdominal tissues, while the malaria parasites are in the mosquito midgut, suggests communication between the midgut epithelial cells and the abdominal tissue which has not yet had direct contact with the parasites. Free radical production in mosquito midgut and haemolymph during Plasmodium infection and their inductive effect on AMPs in abdominal tissues indicates the possible participation of these radicals in mediating a systemic immune response in this mosquito.
Copyright © 2010 Elsevier Ltd. All rights reserved.PMID: 20708028 [PubMed - in process]
Gene. 2011 Jan 1;470(1-2):46-52. Epub 2010 Oct 14.
Functional conservation and divergence of duplicated insulin-like growth factor 2 genes in grass carp (Ctenopharyngodon idellus).
Yuan XN, Jiang XY, Pu JW, Li ZR, Zou SM.
Key Laboratory of Aquatic Genetic Resources and Utilization, Shanghai Ocean University, Huchenghuan Road 999, Shanghai 201306, China.
Abstract
Insulin-like growth factor 2 (IGF2) is a potent mitogenic and survival factor involved in the regulation of growth, development and reproduction in animals. Only one IGF2 gene exists in mammals. Recently, two igf2 genes have been identified in zebrafish, which presumably resulted from gene duplication. However, sequence information of duplicated igf2s and their functional regulation in other teleost fish is still unknown. Here, we report the identification of two igf2 cDNAs in grass carp, Ctenopharyngodon idellus. Like their human ortholog, grass carp igf2a and igf2b mRNAs encoded two structurally distinct mature IGF peptides. Both of them were detected by RT-PCR throughout embryogenesis. Ubiquitous expression of igf2b mRNAs was observed in embryos, whereas igf2a mRNAs were expressed mainly in the notochord and brain with in situ hybridization. In adult fish, igf2b mRNAs were transcribed in multiple tissues, whereas igf2a mRNAs were detected mainly in the liver. Hepatic levels of igf2a and igf2b transcripts were both up-regulated by growth hormone injection. Furthermore, the levels of hepatic igf2a and igf2b mRNAs decreased significantly during starvation and were rebounded rapidly after re-feeding. Our results suggest that duplicated igf2 genes have evolved divergent yet played an overlapping biological role in regulating grass carp growth and development.
Copyright © 2010 Elsevier B.V. All rights reserved.PMID: 20951190 [PubMed - in process]
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