共查询到20条相似文献,搜索用时 15 毫秒
1.
Lenka Monincová Václav Veverka Jiřina Slaninová Miloš Buděšínský Vladimír Fučík Lucie Bednárová Jakub Straka Václav Čeřovský 《Journal of peptide science》2014,20(6):375-384
A novel antimicrobial peptide, designated macropin (MAC‐1) with sequence Gly‐Phe‐Gly‐Met‐Ala‐Leu‐Lys‐Leu‐Leu‐Lys‐Lys‐Val‐Leu‐NH2, was isolated from the venom of the solitary bee Macropis fulvipes. MAC‐1 exhibited antimicrobial activity against both Gram‐positive and Gram‐negative bacteria, antifungal activity, and moderate hemolytic activity against human red blood cells. A series of macropin analogs were prepared to further evaluate the effect of structural alterations on antimicrobial and hemolytic activities and stability in human serum. The antimicrobial activities of several analogs against pathogenic Pseudomonas aeruginosa were significantly increased while their toxicity against human red blood cells was decreased. The activity enhancement is related to the introduction of either l ‐ or d ‐lysine in selected positions. Furthermore, all‐d analog and analogs with d ‐amino acid residues introduced at the N‐terminal part of the peptide chain exhibited better serum stability than did natural macropin. Data obtained by CD spectroscopy suggest a propensity of the peptide to adopt an amphipathic α‐helical secondary structure in the presence of trifluoroethanol or membrane‐mimicking sodium dodecyl sulfate. In addition, the study elucidates the structure–activity relationship for the effect of d ‐amino acid substitutions in MAC‐1 using NMR spectroscopy. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
2.
Misako Taichi Tohimasa Yamazaki Kazuki Kawahara Daisuke Motooka Shota Nakamura Shusaku Harada Tadashi Teshima Tadayasu Ohkubo Yuji Kobayashi Yuji Nishiuchi 《Journal of peptide science》2010,16(7):329-336
A 12‐residue MST isolated from a marine organism is a potent serine protease inhibitor that has a double cyclic structure composed of two ester linkages formed between the β‐hydroxyl and β‐carboxyl groups, Thr3‐Asp9 and Ser8‐Asp11. MST was synthesized by a regioselective esterification procedure employing two sets of orthogonally removable side‐chain protecting groups for the Asp and Ser/Thr residues. In the MST molecule, there were no significant changes observed in yield by changing the order of esterification. SAR study of MST revealed that the minimum required structure for expressing the inhibitory activity is the sequence (1–9) in a monocyclic structure where Pro7 located in the ring plays a crucial role in keeping the structural rigidity. By applying the structural motif of MST, we rationally designed protease inhibitory specificities that differ from those of the natural product. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
3.
S. Omar M. Lalonde M. Marcotte M. Cook J. Proulx K. Goel T. Durst B. J. R. Philogne J. T. Arnason 《Agricultural and Forest Entomology》2000,2(4):253-257
1 Thirty extracts of wood and bark of hardwood trees from Eastern North America were examined for insect growth‐reducing activity in a bioassay with European corn borer, Ostrinia nubilalis, and an antifeedant bioassay with the rice weevil, Sitophilus oryzae. 2 Nine of the bark extracts and four of the wood extracts showed significant growth reducing effects at 0.5% in meridic diets, whereas only two bark extracts and one wood extract showed significant antifeedant effect at the same concentration. 3 Slower growing tree species were more biologically active than fast growing ones. Isolation of the bioactive compounds in one of the active species, Prunus serotina, showed that naringenin, its derivative methoxynaringenin, and eriodictyol were responsible for the antifeedant effects. 相似文献
4.
Borim An Ji-Young Lim Suji Jeong Dong-Mi Shin Eun Young Choi Chang-Ki Min Seok-Ho Hong 《Biochemical and biophysical research communications》2018,495(1):519-525
Myeloid-derived suppressor cells (MDSCs) are a heterogeneous population of cells that regulate immune responses in cancer and various pathological conditions. However, the phenotypic and functional heterogeneity of human MDSCs represents a major hurdle for the development of therapeutic strategies targeting or regulating MDSCs in tumor progression, inflammation, and graft-versus-host disease (GVHD). We previously shown that circulating HLA-DR-CD14+ monocytic MDSCs are a major contributor to clinical outcomes after allogeneic hematopoietic stem cell transplantation (allo-HSCT). In this study, we identified, using high-throughput screening, a set of surface markers that are strongly expressed in HLA-DR-CD14+ monocytic MDSCs isolated from the peripheral blood (PB) of patients receiving allo-HSCT. Subsequent experiments showed the consistent dominant expression of CD1d in monocytic MDSCs of allo-HSCT PB in comparison with granulocytic MDSCs. In addition, CD1d-expressing cells isolated from PB of allo-HSCT patients showed the suppressive activity of T cell proliferation and higher expression of MyD88 and IDO compared with CD1d? cells. Our results suggest that CD1d could be a valuable marker for further therapeutic evaluation of human monocytic MDSCs for immune-related diseases, including GVHD. 相似文献
5.
Setsuko Ando Keitarou Mitsuyasu Yoshitake Soeda Mariko Hidaka Yuki Ito Kouki Matsubara Mitsuno Shindo Yoshiki Uchida Haruhiko Aoyagi 《Journal of peptide science》2010,16(4):171-177
A series of Trp and Arg analogs of antibacterial indolicidin (Ind) was synthesized and the antimicrobial and hemolytic activities were investigated. [L9]Ind, [L11]Ind, [K8,L9]Ind and [K6, 8,L9]Ind showed desirable characteristics, exhibiting negligible hemolytic activity while keeping strong antibacterial activity. The results indicated that the Trp residue at position 11 essentially contributes to both activities and one can not be exchanged for the other, whereas the Trp residues at positions 4 and 9 play important roles in antimicrobial and hemolytic activities, respectively. The Trp residues at positions 6 and 8 play no important roles in biological activities. We then found that the retro analog of Ind showed higher antibacterial activity than Ind against both Gram‐positive and Gram‐negative bacteria but remarkably lower hemolytic activity than that of Ind. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
6.
Optimization of colorectal cancer vaccine candidate protein GA733‐Fc expression in a baculovirus–insect cell system
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The baculovirus–insect cell expression system has been used to produce functional recombinant proteins. The antigen GA733 is a cell‐surface glycoprotein highly expressed on most human colorectal carcinoma cells. Conditions for the expression of GA733 fused to the human immunoglobulin IgG Fc fragment (GA733‐Fc) were optimized in the baculovirus expression system. Several variable factors were adjusted to optimize expression, including the cell line (Sf9 and High Five), multiplicity of infection (MOI) value (0.05, 0.1, 0.5, 1 and 3), post‐infection time (48, 72 and 96 h) and harvested sample (cell culture media (CM) or cell lysate (CL)). In addition, two pFastBac Dual vectors carrying the GA733‐Fc gene were constructed to express GA733‐Fc with or without an endoplasmic reticulum (ER) retention sequence KDEL and used to generate recombinant baculoviruses. Western blot showed that expression depended on the conditions used to express the recombinant proteins. The protein production level and secretion capability differed in each cell line. In Sf9 cells, the highest expression in the CM and CL was obtained with GA733‐Fc at 96 h post‐infection at 0.1 MOI and with GA733‐FcK at 96 h post‐infection at 3 MOI, respectively. In High Five cells, the highest expression in the CM and CL was obtained with GA733‐Fc at 48 h post‐infection at 1 MOI and with GA733‐FcK at 48 h post‐infection at 3 MOI, respectively. These results suggest that the MOI value, post‐infection time and subcellular localization affect expression, and that these conditions can be modified to optimize protein expression in the baculovirus–insect cell system. 相似文献
7.
Daria Leali Roberta Bianchi Antonella Bugatti Stefania Nicoli Stefania Mitola Laura Ragona Simona Tomaselli Grazia Gallo Sergio Catello Vincenzo Rivieccio Lucia Zetta Marco Presta 《Journal of cellular and molecular medicine》2010,14(8):2109-2121
Fibroblast growth factor‐2 (FGF2) plays a major role in angiogenesis. The pattern recognition receptor long‐pentraxin 3 (PTX3) inhibits the angiogenic activity of FGF2. To identify novel FGF2‐antagonistic peptide(s), four acetylated (Ac) synthetic peptides overlapping the FGF2‐binding region PTX3‐(97–110) were assessed for their FGF2‐binding capacity. Among them, the shortest pentapeptide Ac‐ARPCA‐NH2 (PTX3‐[100–104]) inhibits the interaction of FGF2 with PTX3 immobilized to a BIAcore sensorchip and suppresses FGF2‐dependent proliferation in endothelial cells, without affecting the activity of unrelated mitogens. Also, Ac‐ARPCA‐NH2 inhibits angiogenesis triggered by FGF2 or by tumorigenic FGF2‐overexpressing murine endothelial cells in chick and zebrafish embryos, respectively. Accordingly, the peptide hampers the binding of FGF2 to Chinese Hamster ovary cells overexpressing the tyrosine‐kinase FGF receptor‐1 (FGFR1) and to recombinant FGFR1 immobilized to a BIAcore sensorchip without affecting heparin interaction. In all the assays the mutated Ac‐ARPS A‐NH2 peptide was ineffective. In keeping with the observation that hydrophobic interactions dominate the interface between FGF2 and the FGF‐binding domain of the Ig‐like loop D2 of FGFR1, amino acid substitutions in Ac‐ARPCA‐NH2 and saturation transfer difference‐nuclear magnetic resonance analysis of its mode of interaction with FGF2 implicate the hydrophobic methyl groups of the pentapeptide in FGF2 binding. These results will provide the basis for the design of novel PTX3‐derived anti‐angiogenic FGF2 antagonists. 相似文献
8.
利用Bello Cell这种新型的生物反应器,来培养COS7细胞和昆虫Sf9细胞。COS7细胞和昆虫sf9细胞分别用T75培养瓶及spinnerflask培养之后,经细胞计数接入Bello Cell之中,同时检测并分析培养基中葡萄糖、谷氨酰胺、乳酸和氨浓度变化情况。COS7细胞初始接种量为4.208×10^7cells,最终在培养156h后细胞数量达到了4.68×10^8cells,是初始细胞量的11倍。sf9细胞初始接种细胞量为1×10^8cells,在培养192h时,细胞总量达到了最高为4.01×10^9cells,是最初细胞量的40倍。培养基的代谢物进行有规律的变化。Bello Cell适合COS7细胞和昆虫Sf9细胞高密度大规模培养,为动物细胞高效大规模表达药物蛋白,奠定重要的基础。 相似文献
9.
Tatiana M. Domingues Marcus V. Buri Sirlei Daffre Patricia T. Campana Karin A. Riske Antonio Miranda 《Journal of peptide science》2014,20(6):421-428
Gomesin (Gm) has a broad antimicrobial activity making it of great interest for development of drugs. In this study, we analyzed three Gm analogs, [Trp1]‐Gm, [Trp7]‐Gm, and [Trp9]‐Gm, in an attempt to gain insight into the contributions of different regions of the peptide sequence to its activity. The incorporation of the tryptophan residue in different positions has no effect on the antimicrobial and hemolytic activities of the Gm analogs in relation to Gm. Spectroscopic studies (circular dichroism, fluorescence and absorbance) of Gm and its analogs were performed in the presence of SDS, below and above its critical micelle concentration (CMC) (~8 mM), in order to monitor structural changes induced by the interaction with this anionic surfactant (0–15 mM). Interestingly, we found that the analogs interact more strongly with SDS at low concentrations (0.3‐6.0 mM) than close to or above its CMC. This suggests that SDS monomers are able to cover the whole peptide, forming large detergent‐peptide aggregates. On the other hand, the peptides interact differently with SDS micelles, inserting partially into the micelle core. Among the peptides, Trp in position 1 becomes more motionally‐restricted in the presence of SDS, probably because this residue is located at the N‐terminal region, which presents higher conformational freedom to interact stronger with SDS molecules. Trp residues in positions 7 and 9, close to and in the region of the turn of the molecule, respectively, induced a more constrained structure and the compounds cannot insert deeper into the micelle core or be completely buried by SDS monomers. Copyright © 2014 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
10.
Se Ra Park Kisung Ko Sohee Lim So Yeon Cha Hyun Joo Chung Soon Ju Park Soon‐Chul Myung Mi Kyung Kim 《Entomological Research》2020,50(4):199-204
The monoclonal antibody (mAb) CO17‐1A specifically binds to the tumor‐associated cell surface glycoprotein GA733 in colorectal cancer cells. Thus, mAb CO17‐1A has the potential to act as an immune therapeutic protein against colorectal cancer. Recently, it was shown that the baculovirus insect cell expression system produces anti‐colorectal cancer mAb CO17‐1A. In this study, the colorectal cancer antibody mAb CO17‐1A fused to the endoplasmic reticulum (ER) retention signal sequence (KDEL), and the (mAb CO17‐1AK) was expressed in Spodoptera frugiperda Sf9 insect cells. The yield, cell cytotoxicity, and in vitro anti‐tumor activity of mAb CO17‐1AK were verified. Western blotting was performed to confirm that both heavy and light chains of mAb CO17‐1A were expressed in Sf9 insect cells. The insect‐derived mAb (mAbI) CO17‐1A was purified using a protein G affinity column. An in vitro wound healing assay was conducted to determine the inhibition activity of mAb CO17‐1A during tumor cell migration, showing that mAbI CO17‐1AK was effective as mammalian‐derived mAb CO17‐1A (mAbM CO17‐1A). These results suggest that the insect cell expression system can produce and properly assemble mAbs that inhibit tumor cell migration. 相似文献
11.
Translocation of several fluorescently labeled arginine‐rich peptides into intact plant cells was quantitatively examined in order to investigate the structural factors required for efficient cellular internalization, and thereby, to evaluate the potential of arginine‐rich peptides as intracellular delivery vectors in plants. Cell‐penetrating peptides (CPPs) such as arginine‐rich peptides permit the direct introduction of biologically active macromolecules into plant cytoplasm to manipulate various intracellular processes. While a significant level of adsorption of applied arginine‐rich peptides was observed in the cell walls rich in negative charges, removal of adsorbed peptides by trypsin treatment allowed determination of the amount of internalized peptides in a quantitative manner using spectrofluorometric analysis. The internalization of arginine‐rich peptides depended on the number of arginine residues, and the peptide containing eight arginine residues showed most effective internalization. Besides, the position of small cargoes attached to the arginine‐rich peptides markedly affected the internalization efficiency. The results obtained in this study provide useful information for the development of efficient intracellular delivery tools in plant science. Copyright © 2008 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
12.
Mihály L. Kecskés Michael T. Rose Erwin Michel Benôit Lauby Malala Rakotondrainibe Andrea Casteriano Attila Palágyi Anaëlle Moutouvirin Sébastien Elter Romain Guillas Ganisan Krishnen Ivan R. Kennedy 《Engineering in Life Science》2009,9(6):431-436
Rapid, specific techniques are essential to monitor the quality of inoculant plant growth‐promoting strains at all stages of manufacture from starter culture to the final product in its carrier medium. In this study, colony immunoblotting was evaluated for the specific detection and enumeration of Citrobacter freundii, one component of a Vietnamese commercial inoculant plant growth‐promoting product used to improve the yield and nutrient efficiency of paddy rice. For quality control of either sterilised or unsterilised carrier media in commercial products colony immunoblotting proved to be a promising tool. Furthermore, it was possible using this technique to measure the survival of this strain in soil and the rhizosphere. 相似文献
13.
Screening, isolation and in vitro assays have been used for characterization of antioxidative peptides derived from food proteins, and incompatible deductions of structural characteristics derived from the isolated peptides have been brought forward. However, there is still little information concerning the structure‐activity relationship of antioxidative peptides. QSAR modeling was performed, respectively, on synthetic tripeptides and tetrapeptides related to LLPHH. According to cumulative squared multiple correlation coefficients (R2), cumulative cross‐validation coefficients (Q2) and relative standard deviation for calibration set (RSDc), two credible models for tripeptide and tetrapeptide databases, respectively, have been built with partial least squares (PLS) regression (R2 for models of tripeptide and tetrapeptide are 0.744 and 0.943, Q2 are 0.631 and 0.414, and RSDc are 0.323 and 0.111, respectively). Meanwhile, according to the cumulative multiple correlation coefficient for the predictive set ($R_{\rm {ext}}^{2}$ ) and the relative standard deviation for the predictive set (RSDp), the predictive ability of the model for tripeptides also is excellent ($R_{\rm {ext}}^{2}$ and RSDp are 0.719 and 0.450, respectively). Hydrogen bond property and hydrophilicity of the amino acid residue next to the C‐terminus, and the hydrophobicity as well as electronic propertyof the N‐terminus are more significant; meanwhile, the electronic property of the C‐terminus is beneficial for antioxidant activity. The structural characteristics we found are very useful in understanding and predicting the peptide structures responsible for activity and development of functional foods with peptides as active compounds, or antioxidative peptides as alternatives to other antioxidants. Copyright © 2011 European Peptide Society and John Wiley & Sons, Ltd. 相似文献
14.
15.
Jennifer M. Sealy Anh P. Truong Luke Tso Gary D. Probst Jose Aquino Roy K. Hom Barbara M. Jagodzinska Darren Dressen David W.G. Wone Louis Brogley Varghese John Jay S. Tung Michael A. Pleiss John A. Tucker Andrei W. Konradi Michael S. Dappen Gergely Toth Hu Pan Lany Ruslim Jim Miller Michael P. Bova Sukanto Sinha Kevin P. Quinn John-Michael Sauer 《Bioorganic & medicinal chemistry letters》2009,19(22):6386-6391
Using structure-guided design, hydroxyethylamine BACE-1 inhibitors were optimized to nanomolar Aβ cellular inhibition with selectivity against cathepsin-D. X-ray crystallography illuminated the S1′ residues critical to this effort, which culminated in compounds 56 and 57 that exhibited potency and selectivity but poor permeability and high P-gp efflux. 相似文献
16.
17.
Jeong‐Hwan Lee Kyung‐A Hwang SungSu Park Young‐Kug Choo Kisung Ko 《Entomological Research》2014,44(5):207-214
The anti‐breast cancer monoclonal antibody (mAb) BR55 was expressed in the baculovirus–insect cell expression system, which is advantageous because of its high production capacity, cell culture flexibility and glycosylation capability. The baculovirus–insect cell expression system was successfully established for production of mAb BR55 and mAb BR55 fused with the KDEL (Lys–Asp–Glu–Leu) endoplasmic reticulum (ER) retention signal (mAb BR55K). The heavy chain (HC) and light chain (LC) genes of mAb BR55 were cloned under the control of the polyhedrin (PPH) and P10 promoters, respectively, in the pFastBacDual vector. The antibody gene‐expression cassettes carrying both the HC and LC genes were transferred into a bacmid in Escherichia coli (DH10Bac). The bacmid carrying the expression cassettes was transfected into Sf9 insect cells to generate baculovirus expressing mAb BR55 and BR55K. Western blot analysis confirmed the expression of mAb BR55 and BR55K in baculovirus‐infected insect cells. Cell direct enzyme linked immunosorbent assay (ELISA) showed that both mAbs from insect cell lysates or cell culture medium bound to MCF‐7 human breast cancer cells. Both mAb BR55 and BR55K were successfully purified using a Protein A affinity column. Collectively, these results suggest that the anti‐breast cancer mAb BR55 can be expressed, properly assembled and purified from the baculovirus expression system, which can serve as an alternative system for antibody production. 相似文献
18.
A. Contreras‐Gómez A. Sánchez‐Mirón F. García‐Camacho E. Molina‐Grima Y. Chisti 《Biotechnology progress》2014,30(1):1-18
The baculovirus‐insect cell expression system is widely used in producing recombinant proteins. This review is focused on the use of this expression system in developing bioprocesses for producing proteins of interest. The issues addressed include: the baculovirus biology and genetic manipulation to improve protein expression and quality; the suppression of proteolysis associated with the viral enzymes; the engineering of the insect cell lines for improved capability in glycosylation and folding of the expressed proteins; the impact of baculovirus on the host cell and its implications for protein production; the effects of the growth medium on metabolism of the host cell; the bioreactors and the associated operational aspects; and downstream processing of the product. All these factors strongly affect the production of recombinant proteins. The current state of knowledge is reviewed. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 30:1–18, 2014 相似文献
19.
Christian D. Taeger Wibke Müller‐Seubert Raymund E. Horch Konstantin Präbst Frank Münch Carol I. Geppert Torsten Birkholz Adrian Dragu 《Journal of cellular and molecular medicine》2014,18(5):885-894
Tissue undergoing free transfer in transplant or reconstructive surgery always is at high risk of ischaemia‐related cell damage. This study aims at assessing different procedures using an extracorporeal perfusion and oxygenation system to investigate the expression of hypoxia inducible factor (HIF)‐1‐α as marker for hypoxia and of the pro‐apoptotic protein Caspase‐3 in skeletal muscle to elucidate potential improvements in tissue conservation. Twenty‐four porcine rectus abdominis muscles were assigned to five different groups and examined after they had been extracorporeally preserved for 60 min. time. Group I was left untreated (control), group II was perfused with a cardioplegic solution, group III was flushed with 10 ml of a cardioplegic solution and then left untreated. Group IV and V were perfused and oxygenated with either an isotone crystalloid solution or a cardioplegic solution. Among others, immunohistochemistry (Caspase‐3 and HIF‐1‐α) of muscle samples was performed. Furthermore, oxygen partial pressure in the perfusate at the arterial and venous branch was measured. Expression of Caspase‐3 after 60 min. was reduced in all groups compared to the control group. Furthermore, all groups (except group III) expressed less HIF‐1‐α than the control group. Oxygenation leads to higher oxygen levels at the venous branch compared to groups without oxygenation. Using an extracorporeal perfusion and oxygenation system cell damage could be reduced as indicated by stabilized expressions of Caspase‐3 and HIF‐1‐α for 60 min. of tissue preservation. Complete depletion of oxygen at the venous branch can be prevented by oxygenation of the perfusate with ambient air. 相似文献
20.
Rasha M. Hassan Ali M. Yehia Ola A. Saleh Aida A. El‐Azzouny Hassan Y. Aboul‐Enein 《Chirality》2018,30(6):828-836
Fluoroquinolones are popular class of antibiotics with distinct chemical functionality. Most of them are ampholytes with one chiral center. Stereogeneic center is located either in the side ring of Gatifloxacin (GFLX) or in the quinolone core of Ofloxacin (OFLX). These two amphoteric fluoroquinolones have terminal amino groups in common. The unusual Nadifloxacin (NFLX) is an acidic fluoroquinolone with a core chiral center. Owing to chirality and functionality differences among GFLX, OFLX, and NFLX, we mapped these enantiomers onto structure‐retention relationship. Amount of acetic acid modifier was studied in screened mobile phase and cellulose tris(3‐chloro‐4‐methyl phenyl carbamate) (Lux cellulose‐2) stationary phase. Experimental design of acetic acid% along with column temperature have been applied. Resolution and enantioselectivity have been related to structural features of the studied enantiomers. High amount of acid (0.4%) was optimum for the separation of either side chirality with a proximate amino group (GFLX) or core chirality without basic functionality (NFLX), while low amount (0.2%) is optimum for core chiral center with distal amino group (OFLX). Temperature has no significant effect on resolution and retention of enantiomers except for OFLX. Enantio‐retention explains possible chiral selective and nonselective interactions. The proposed methods have been validated for pharmaceutical analyses. 相似文献