摘要:

The morphology of structures formed by the self‐assembly of short N‐terminalt‐butyloxycarbonyl (Boc) and C‐terminal methyl ester (OMe) protected and Boc‐deprotected hydrophobic peptide esters was investigated. We have observed that Boc‐protected peptide esters composed of either only aliphatic hydrophobic amino acids or aliphatic hydrophobic amino acids in combination with aromatic amino acids, formed highly organized structures, when dried from methanol solutions. Transmission and scanning electron microscopic images of the peptides Boc‐Ile‐Ile‐OMe, Boc‐Phe‐Phe‐Phe‐Ile‐Ile‐OMe and Boc‐Trp‐Ile‐Ile‐OMe showed nanotubular structures. Removal of the Boc group resulted in disruption of the ability to form tubular structures though spherical aggregates were formed. Both Boc‐Leu‐Ile‐Ile‐OMe and H‐Leu‐Ile‐Ile‐OMe formed only spherical nanostructures. Dynamic light scattering studies showed that aggregates of varying dimensions were present in solution suggesting that self‐assembly into ordered structures is facilitated by aggregation in solution. Fourier transform infrared spectroscopy and circular dichroism spectroscopy data show that although all four of the protected peptides adopt well‐defined tertiary structures, upon removal of the Boc group, only H‐Phe‐Phe‐Phe‐Ile‐Ile‐OMe had the ability to adoptβ‐structure. Our results indicate that hydrophobic interaction is a very important determinant for self‐assembly and presence of charged and aromatic amino acids in a peptide is not necessary for self‐

ISSN:1075-2617    

DOI:10.1002/psc.2395

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

Successful and effective cellular delivery remains a main obstacles in the medical field. The use of cell‐penetrating peptides (CPPs) has become one of the most important tools for the internalisation of a wide range of molecules including pharmaceuticals. It is still difficult to choose one CPP for one biological application because there is no ubiquitous CPP meeting the diverse requirements. In our case, we are looking for a suitable CPP to deliver the pro‐apoptotic KLA peptide (KLAKLAKKLAKLAK) by a simple co‐incubation strategy. For that reason, we selected three different cell lines (fibroblastic, cancerous and macrophagic cells) and studied the uptake and subcellular localisation of six different CPPs alone as well as mixed with the KLA peptide. Furthermore, we used the CPPs with a carboxyamidated or a carboxylated C‐terminus and analysed the impact of the C‐termini on internalisation and cargo delivery. We could clearly showed that the cellular CPP uptake is not only dependent on the used CPP and cell line but also highly affected by its chemical nature of the C‐terminus (uptake: carboxyamidated CPPs > carboxylated CPPs) and can influence its cellular localisation. We successfully delivered the KLA peptide in the three cell lines and learned that here as well, the C‐terminus is crucial for an effective peptide delivery. Finally, we induced apoptosis in mouse leukaemic monocyte macrophage (RAW 264.7) and in human breast adenocarcinoma (MCF‐7) cells using the mixture of amidated MPG peptide : KLA and in african green monkey kidney fibroblast (Cos‐7) cells using carboxylated integrin peptide : KLA. Copyright © 2012 European Peptide So

ISSN:1075-2617    

DOI:10.1002/psc.2396

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

We have been engaged in the microwave‐solid phase peptide synthesis (SPPS) synthesis of the phenylglycine (Phg)‐containing pentapeptide H‐Ala‐Val‐Pro‐Phg‐Tyr‐NH2(1) previously demonstrated to bind to the so‐called BIR3 domain of the anti‐apoptotic protein XIAP. Analysis of the target peptide by a combination of RP‐HPLC, ESI‐MS, and NMR revealed the presence of two diastereoisomers arising out of the racemisation of the Phg residue, with the percentage of the LLLDL component assessed as 49%. We performed the synthesis of peptide (1) using different microwave and conventional stepwise SPPS conditions in attempts to reduce the level of racemisation of the Phg residue and to determine at which part of the synthetic cycle the epimerization had occurred. We determined that racemisation occurred mainly during the Fmoc‐group removal and, to a much lesser extent, during activation/coupling of the Fmoc‐Phg‐OH residue. We were able to obtain the desired peptide with a 71% diastereomeric purity (29% LLLDL as impurity) by utilizing microwave‐assisted SPPS at 50 °C and power 22 Watts, when the triazine‐derived coupling reagent DMTMM‐BF4was used, together with NMM as an activator base, for the incorporation of this residue and 20% piperidine as an Fmoc‐deprotection base. In contrast, the phenylalanine analogue of the above peptide, H‐Ala‐Val‐Pro‐Phe‐Tyr‐NH2(2), was always obtained as a single diastereoisomer by using a range of standard coupling and deprotection conditions. Our findings suggest that the racemisation of Fmoc‐Phg‐OH, under both microwave‐SPPS and stepwise conventional SPPS syntheses conditions, is very facile but can be limited through the use of the above stated conditions. Copyri

ISSN:1075-2617    

DOI:10.1002/psc.2398

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

Native chemical ligation of unprotected peptides in organic solvents has been previously reported as a fast, efficient, and suitable method for coupling of hydrophobic peptides. However, it has not been determined whether the reaction can be carried out without possible side reactions or racemization. Here, we present a study on the chemoselectivity of this method by model reactions designed to test the reactivity of Arg and Lys side chains as well as that ofα‐amino groups. A possible racemization of the C‐terminal amino acid of the N‐terminal peptide was also investigated. The results show that ligation in organic solvents can be conducted chemoselectively without side reactions with other nucleophilic groups. Furthermore, no racemization of the C‐terminal amino acid was observed if both educts were added simultaneously. Thus, native chemical ligation can be performed either in aqueous buffer systems or in organic solvents paving the way for the synthesis of larger hydrophobic peptides and/or membrane proteins. Copyright © 2012 European Peptide Society and John Wiley&

ISSN:1075-2617    

DOI:10.1002/psc.2401

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

We utilised a simple bidirectional (N→C and C→N) solid phase synthesis strategy entailing conventional solid phase peptide synthesis and fragment condensation with a water‐soluble carbodiimide to synthesise a model anionic glycylglycine bolaamphiphile containing a suberic acid linker moiety, namelyN,N′‐suberoyldiglycylglycine. The synthetic suberoyldiglycylglycine was purified using its inherent ability to rapidly self‐assemble in an aqueous acidic solution (0.1% trifluoroacetic acid). Monitoring of the rapid assembly process corroborated our visual observation and confirmed packing‐directed self‐assembly rather than non‐specific aggregation or precipitation. The progress of suberoyldiglycylglycine self‐assembly was observed to be via the formation of oligomers in the solution, which then self‐assembled to form layeredβ‐sheet type macrostructures. Within 24 h, nanotubes grew from these macrostructures and eventually combined to formed microtubes, which we isolated after 5–7 days. Copyright © 2012 European Peptide S

ISSN:1075-2617    

DOI:10.1002/psc.2402

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

Synthetic peptides derived from GB virus C (GBV‐C) have previously been studied in our group for the development of new systems capable of diagnosing diseases caused by this humanotropic virus. We also recently described specific peptide domains of the E2 envelop protein of GBV‐C that have the capacity to interfere with the HIV‐1 fusion peptide, produce a notable decrease in cellular membrane fusion, and perturb HIV‐1 infectivity in a dose‐dependent manner.The present work discloses the design and synthesis of both linear and cyclic branched peptides based on a previously reported N‐terminal sequence of the GBV‐C E2 protein. Immunoassays and cell–cell fusion assays were performed to evaluate their diagnostic value to detect anti‐GBV‐C antibodies in HIV‐1 patients, as well as their putative anti‐HIV‐1 activity as entry inhibitors.Our results showed that chemical modifications of the selected E2(7–26) linear peptide to afford cyclic architecture do not result in an enhanced inhibition of gp41 HIV‐1‐mediated cell–cell fusion nor improved sensitivity in the detection of GBV‐C antibodies in HIV‐1 co‐infected patients. Thus, the ELISA data reinforce the potential utility of linear versions of the E2(7–26) region for the development of new peptide‐based immunosensor devices for the detection of anti‐GBV‐C antibodies in HIV‐1 co‐infected patients. Copyright © 20

ISSN:1075-2617    

DOI:10.1002/psc.2403

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

Three fluorescein derivatives of human insulin (HI,1) labeled at positionsNαA1,NαB1andNεB29respectively, were synthesized using anN‐trifluoroacetyl‐based protecting group scheme. The Tfa protecting group introduced by reaction with ethyl trifluoroacetate was found to be stable in aqueous and organic media and efficiently removed under mild basic conditions. Copyright © 2012 European Peptide Society and John Wiley&So

ISSN:1075-2617    

DOI:10.1002/psc.2405

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

Light color and savory flavor enhancer are attractive for consumers and food producers. The effect of addition time ofl‐cysteine on inhibiting color formation was investigated in soybean peptide‐xylose system, and the possible pathway was explored. Once dicarbonyl compounds were formed during the Maillard reaction, the addition ofl‐cysteine had no color‐inhibiting effect; ifl‐cysteine was added immediately after the Amadori compound was formed, the extraordinary color‐inhibiting effect was observed. Therefore, an improved way to inhibit color formation was proposed on the basis of the interaction ofl‐cysteine and Amadori compounds by controlling the addition time ofl‐cysteine through gradient temperature‐elevating Maillard reaction. The system was heated at 80 °C for 60 min to form Amadori compounds, followed by the addition of L‐cysteine, and the temperature was raised to 120 °C and held for 110 min. Compared with traditional products, the lightest color product was found desirable by GC/MS analysis and sensory evaluation. The novel method proposed can be a guide for the industrial preparation of light‐colored products. Copyright © 2012 European Peptide

ISSN:1075-2617    

DOI:10.1002/psc.2406

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY

摘要:

Transglutaminase 2 (TG2) is an autoantigen in celiac disease (CD) and it has multiple biologic functions including involvement in cell adhesion through interactions with integrins, fibronectin (FN), and heparan sulfate proteoglycans. We aimed to delineate the heparin‐binding regions of human TG2 by studying binding kinetics of the predicted heparin‐binding peptides using surface plasmon resonance method. In addition, we characterized immunogenicity of the TG2 peptides and their effect on cell adhesion. The high‐affinity binding of human TG2 to the immobilized heparin was observed, and two TG2 peptides, P1 (amino acids 202–215) and P2 (261–274), were found to bind heparin. The amino acid sequences corresponding to the heparin‐binding peptides were located close to each other on the surface of the TG2 molecule as part of theα‐helical structures. The heparin‐binding peptides displayed increased immunoreactivity against serum IgA of CD patients compared with other TG2 peptides. The cell adhesion reducing effect of the peptide P2 was revealed in Caco‐2 intestinal epithelial cell attachment to the FN and FN‐TG2 coated surfaces. We propose that TG2 amino acid sequences 202–215 and 261–274 could be involved in binding of TG2 to cell surface heparan sulfates. High immunoreactivity of the corresponding heparin‐binding peptides of TG2 with CD patient's IgA supports the previously described role of anti‐TG2 autoantibodies interfering with this interaction. Copyright © 2012 European Peptide Soc

ISSN:1075-2617    

DOI:10.1002/psc.2413

出版商:John Wiley&Sons, Ltd    

年代:2012

数据来源: WILEY