1.Identification of the target site of antimicrobial peptide AMP-17 against Candida albicans.
Longbing YANG ; Zhuqing TIAN ; Luoxiong ZHOU ; Chaoqin SUN ; Mingjiao HUANG ; Chunren TIAN ; Jian PENG ; Guo GUO
Chinese Journal of Biotechnology 2023;39(1):304-317
Candida albicans is one of the major causes of invasive fungal infections and a serious opportunistic pathogen in immunocompromised individuals. The antimicrobial peptide AMP-17 has prominent anti-Candida activity, and proteomic analysis revealed significant differences in the expression of cell wall (XOG1) and oxidative stress (SRR1) genes upon the action of AMP-17 on C. albicans, suggesting that AMP-17 may exert anti-C. albicans effects by affecting the expression of XOG1 and SRR1 genes. To further investigate whether XOG1 and SRR1 genes were the targets of AMP-17, C. albicans xog1Δ/Δ and srr1Δ/Δ mutants were constructed using the clustered regulatory interspaced short palindromic repeats-associated protein 9 (CRISPR/Cas9) system. Phenotypic observations revealed that deletion of two genes had no significant effect on C. albicans growth and biofilm formation, whereas XOG1 gene deletion affected in vitro stress response and mycelium formation of C. albicans. Drug sensitivity assay showed that the MIC80 values of AMP-17 against xog1Δ/Δ and srr1Δ/Δ mutants increased from 8 μg/mL (for the wild type C. albicans SC5314) to 16 μg/mL, while the MIC80 values against srr1Δ/Δ: : srr1 revertants decreased to the level of the wild type SC5314. In addition, the ability of AMP-17 to inhibit biofilm formation of both deletion strains was significantly reduced compared to that of wild type SC5314, indicating that the susceptibility of the deletion mutants to AMP-17 was reduced in both the yeast state and during biofilm formation. These results suggest that XOG1 and SRR1 genes are likely two of the potential targets for AMP-17 to exert anti-C. albicans effects, which may facilitate further exploration of the antibacterial mechanism of novel peptide antifungal drugs.
Humans
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Candida albicans
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Antimicrobial Peptides
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Proteomics
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Peptides/pharmacology*
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Transcription Factors/metabolism*
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Antifungal Agents/pharmacology*
2.Advances in research on cytokine mimetic peptides/non-peptides stimulating hematopoiesis.
Zhi-Hong YUAN ; Yong-Zhi XI ; Fan-Hua KONG
Journal of Experimental Hematology 2002;10(1):73-76
Cytokines such as erythropoietin (EPO) and thrombopoitein (TPO) and so on, which stimulate hematopoiesis, can regulate self-renewal, proliferation, differentiation, maturation and programmed cell death of hematopoietic cells through specifically binding to surface receptors. Recently random phage display peptide libraries and other screening methods have been used to isolate mimetic including small peptides and non-peptides molecules, which can mimic the same effects as cytokines, such as EPO and TPO, and demonstrate the similar potency and activity as EPO and TPO in a panel of in vitro biological assays and in animal experiments. These approaches are critical to further research of interactive mechanisms between cytokine and receptor, receptor activation and rational design of other desired cytokine mimetic. This review concisely introduced recent advances in research on mimetic of EPO, TPO and other cytokines and future directions.
Animals
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Cytokines
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pharmacology
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Erythropoietin
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pharmacology
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Hematopoiesis
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drug effects
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physiology
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Humans
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Peptide Library
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Peptides
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pharmacology
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Thrombopoietin
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pharmacology
3.Progress on the design and optimization of antimicrobial peptides.
Ruonan ZHANG ; Di WU ; Yitian GAO
Journal of Biomedical Engineering 2022;39(6):1247-1253
Antimicrobial peptides (AMPs) are a class of peptides widely existing in nature with broad-spectrum antimicrobial activity. It is considered as a new alternative to traditional antibiotics because of its unique mechanism of antimicrobial activity. The development and application of natural AMPs are limited due to their drawbacks such as low antimicrobial activity and unstable metabolism. Therefore, the design and optimization of derived peptides based on natural antimicrobial peptides have become recent research hotspots. In this paper, we focus on ribosomal AMPs and summarize the design and optimization strategies of some related derived peptides, which include reasonable primary structure modification, cyclization strategy and computer-aided strategy. We expect to provide ideas for the design and optimization of antimicrobial peptides and the development of anti-infective drugs through analysis and summary in this paper.
Antimicrobial Cationic Peptides/chemistry*
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Antimicrobial Peptides
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Drug Design
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Anti-Infective Agents/pharmacology*
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Anti-Bacterial Agents
4.Research progress of natural collagen peptides and its skincare efficacy.
Yaqi WU ; Haiyan JU ; Yonggang LYU
Journal of Biomedical Engineering 2022;39(6):1254-1262
Natural collagen peptides are collagen hydrolysates. Because of their unique physicochemical properties and excellent biological activities, collagen peptides have been a research hotspot of cosmetic raw materials development and skincare efficacy improvement. Combined with the needs of the skincare efficacy and the development trends of cosmetics, the extraction methods and their structural characteristics of natural collagen peptides were summarized in detail. The applications and its research progress in skincare efficacy of collagen peptides, such as moisturizing and anti-wrinkle, trophism and anti-aging, filling and skin regeneration were expressed with emphasis. Finally, the development and practical applications in cosmetics of natural collagen peptides were adequately prospected.
Skin Care
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Skin
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Peptides/pharmacology*
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Cosmetics/chemistry*
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Collagen
5.Advances in several important antimicrobial lipopeptids from Bacillus spp.
Daoming LI ; Ying WANG ; Chao CHEN ; Mingbai ZENG ; Qianru LI ; Qingyun JIA ; Xiuli LIU ; Yongyue HOU ; Chengming FAN ; Yuhong CHEN ; Zanmin HU
Chinese Journal of Biotechnology 2022;38(5):1768-1783
Bacillus spp. are probiotics and can secrete a variety of natural antimicrobiol active substances, of which lipopeptides are an important class. Up to now, about 90 lipopeptides have been identified, and most of them are cyclic lipopeptides. surfactin, iturin, fengycin, bacillomycin and polymyxins are widely studied, and the first three have huge potential for application due to their properties of surfactants and anti-fungal, anti-bacterial, anti-viral, anti-tumor and anti-inflammatory functions. In this paper, the research progress in the structure, function, synthesis regulation, separation, purification and production of surfactin, iturin and fengycin was reviewed. Synthetic biology is a vital means to increase the yield of lipopeptides, and in the future, lipopeptides can be used in crop cultivation, animal farming, food, medicine and petroleum industries as well as environmental protection. Future research should be strengthened on the discovery of new lipopeptides, synthesis of high-activity lipopeptides, economical production of lipopeptides on a large scale and their safety evaluation.
Anti-Bacterial Agents
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Anti-Infective Agents/pharmacology*
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Bacillus
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Bacillus subtilis
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Lipopeptides/pharmacology*
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Peptides, Cyclic/pharmacology*
6.Effects of humanin on elevation of intracellular calcium concentration induced by beta-amyloid peptide(31-35) in cultured cortical neurons..
Ling-Min LI ; Jian-Tian QIAO ; Ce ZHANG
Acta Physiologica Sinica 2009;61(2):127-131
The disruption of the intracellular Ca(2+) homeostasis has been reported to be one of the mechanisms of beta-amyloid (Abeta) neurotoxicity in Alzheimeros disease (AD). Abeta(31-35), a small active fragment of Abeta, is believed to possess the similar biological activities of full-length Abeta molecule. Humanin (HN) is a recently identified peptide that suppresses neuronal death initiated by AD-related insults. The present study was to investigate the effects of HN on Abeta(31-35)-induced elevation of [Ca(2+)](i) in cultured cortical neurons by real-time fluorescence imaging technique using the Ca(2+)-sensitive dye, Fura-2/AM. The elevation of [Ca(2+)](i) was observed in cultured neurons exposed to Abeta(31-35) (25 mumol/L) (F340/F380: 1 042.56+/- 83.54, compared with control group: 804.73+/- 48.230, P<0.05, n=10). Pretreatment of HN (10 mumol/L) for 10 min significantly decreased the elevation of [Ca(2+)](i) induced by Abeta(31-35) (25 mumol/L) (F340/F380: 918.788+/- 50.73, compared with Abeta(31-35) group, P<0.05, n=10). When neurons were treated with HN and Abeta(31-35) simultaneously, HN (10 mumol/L) could not change the elevation of [Ca(2+)](i) induced by Abeta(31-35) (F340/F380: 1 036.68+/- 88.96, compared with Abeta(31-35) group, P>0.05, n=10), while HN (20 mumol/L) diminished the elevation of [Ca(2+)](i) induced by Abeta(31-35) (25 mumol/L) significantly (F340/F380: 898.56+/- 76.46, compared with Abeta(31-35) group, P<0.05, n=10). The findings imply that: (1) the disruption of the calcium homeostasis induced by Abeta(31-35) is possibly the basis of the neurotoxicity of Abeta(31-35) in cultured cortical neurons; (2) HN suppresses the elevation of [Ca(2+)](i) induced by Abeta(31-35) in a dose- and time-dependent manner.
Amyloid beta-Peptides
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pharmacology
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Calcium
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metabolism
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Cell Death
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Cells, Cultured
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Homeostasis
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Humans
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Intracellular Signaling Peptides and Proteins
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pharmacology
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Neurons
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drug effects
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Peptide Fragments
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pharmacology
7.Screening of peptide inhibitors of acetylcholinesterase from 12-mer random phage display peptide library.
Xing-mei ZHANG ; Yu-sheng SHI ; Chun-xia WANG
Journal of Southern Medical University 2006;26(7):1053-1054
OBJECTIVETo screen the peptide inhibitor of acetylcholinesterase (AChE) from 12-mer random phage display peptide library.
METHODSHuman AChE was used as the target to screen its binding peptides from 12-mer random phage display peptide library. The positive phage clones were isolated after three rounds of biopanning followed then by sequence analysis and their activity evaluation.
RESULTSSix positive phage clones binding to human AChE were obtained, and 4 of them sharing the conservative sequence W(S/P)HY inhibited the enzyme activity of AChE.
CONCLUSIONAcquisition of AChE inhibitor from phage display library provides clues for designing peptide inhibitors of AChE.
Acetylcholinesterase ; metabolism ; Cholinesterase Inhibitors ; metabolism ; pharmacology ; Humans ; Peptide Library ; Peptides ; metabolism ; pharmacology ; Protein Binding
8.The protection of scorpion venom derived activity peptide against the change of dynorphin in the early Parkinson's disease rats.
Dong-mei WANG ; Dan ZHAO ; Sheng-ming YIN ; Dong AN ; Wei CHEN ; De-qin YU ; Hong XU ; Jie ZHAO ; Wan-qin ZHANG ; Yu-xiang TIAN
Chinese Journal of Applied Physiology 2015;31(2):120-122
Animals
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Dynorphins
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metabolism
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Parkinson Disease
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drug therapy
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metabolism
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Peptides
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pharmacology
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Rats
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Scorpion Venoms
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pharmacology
9.Anti-tumor efficacy of P53 with 9R cell-penetrating peptides.
Yuan LIU ; Rui CHEN ; Nan ZHANG ; Xianlong YE ; Yin BAI ; Yuquan WEI ; Guiping REN ; Deshan LI
Chinese Journal of Biotechnology 2013;29(7):955-964
To enhance the penetration of P53 into tumor cells by fusion it with the cell penetrating peptide 9R. The fusion gene of 9R-p53 was cloned into the expression vector. The fusion protein, CPPs-P53, was expressed and purified. We detected the rate of cell growth inhibition and apoptosis by MTT and Annexin-V-FITC/PI double stained method respectively for measuring its effect on tumor cells. CPPs-P53 and P53 were successfully expressed and purified, the purity of both proteins reached up to 90%. MTT assay showed that the cell growth inhibition by CPPs-P53 was more efficient than P53, and the rate of cell growth inhibition is dose-dependent. The apoptosis experiment showed that P53 could induce apoptosis of tumor cells. Compared with the P53, CPPs-P53 had a more significant effect in inducing cell apoptosis (**P < 0.01). The CPPs-P53 shows more significant effects than P53 in inhibiting cell growth and inducing apoptosis on tumor cells.
Apoptosis
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Cell Line, Tumor
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Cell-Penetrating Peptides
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pharmacology
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Humans
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Tumor Suppressor Protein p53
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pharmacology
10.Binding characteristics between RGD-containing cyclic peptide and rat hepatic stellate cells: an in vitro study.
Shi-lin DU ; Ji-yao WANG ; Wei-yue LU
Chinese Journal of Hepatology 2005;13(5):362-365
OBJECTIVETo investigate the binding characteristics between an artificial Arg-Gly-Asp (RGD)-containing cyclic peptide [cyclo(CGRGDSPK)] and rat hepatic stellate cells (HSC).
METHODSAn artificial RGD-containing cyclic peptide was labeled with fluorescein isothiocyanate (FITC). HSCs were isolated by collagenase in situ liver recirculating and purified by density gradient centrifugation from normal rats. The cells were cultured for 5 days of primary culture (quiescent phenotype) or for 7 days of secondary culture (activated phenotype). To access the binding and uptake, HSCs were incubated with FITC-cRGD of different concentrations at 4 degree C or 37 degree C, and then the binding and uptake were investigated by flow cytometry. The location of FITC-cRGD in HSC was investigated by fluorescent microscopy. Kd and maximal binding sites per cell were calculated by radioligand binding assay (RBA) of receptors using 3H-cRGD. In the interim, FITC-cAGA was used as a peptide control devoid of any binding site.
RESULTSThe binding between FITC-cRGD and HSC was saturable, time- and dose-dependent and could compete with overdosed unlabeled cRGD. The fluorescence was mainly distributed in cytoplasma, especially near the nuclei. Kd was 7.05 x 10(-9) mol/L and Bmax per cell was nearly 6.79 x 10(5).
CONCLUSIONSThe results demonstrate that cRGD are specifically taken up by HSC through a receptor-mediated pathway. The information is useful for understanding the ligand-receptor interaction of HSC. FITC labeled cyclic RGD-peptides meet the standards of special ligands and FITC does not change the binding activation of cyclic RGD-peptides.
Animals ; Binding Sites ; Cells, Cultured ; Hepatocytes ; cytology ; metabolism ; Oligopeptides ; pharmacology ; Peptides, Cyclic ; pharmacology ; Protein Binding ; Rats