Celimar Sinézia, Tháyna Sisnande, Luis Peña Icart, Luís Maurício T. R. Lima
Amylin receptor agonism safely benefit diabetic patients, reducing the insulin requirements and glycemic excursions. Pramlintide is the triple proline human amylin analogue first used as injectable drug, but lacking physico-chemical compatibility when co-formulated with insulin. Here, we report the design and characterization of polymeric microparticles for oral delivery of pramlintide. Eudragit S100, a gastric-resistant polymer, was used in preparation of pramlintide-loaded spherical microcapsules by double emulsion and solvent evaporation technique, with approximately 66 μm ± 11 particle size, with 83.2% ± 2.7 efficiency for pramlintide entrapment and 67.6% ± 2.1 yield. Intra-venous pramlintide free in solution showed a plasmatic half-life of 6.8 min in mice. In contrast, oral delivery of acid-resistant pramlintide-loaded microparticles in mice showed a protracted release for 120 min compared to 30 min obtained for pramlintide in solution. Our data provide evidences for the potential use of the oral route in the therapeutic development of pramlintide formulations.
{"title":"Oral delivery of the amylin receptor agonist pramlintide","authors":"Celimar Sinézia, Tháyna Sisnande, Luis Peña Icart, Luís Maurício T. R. Lima","doi":"10.1002/pep2.24346","DOIUrl":"https://doi.org/10.1002/pep2.24346","url":null,"abstract":"Amylin receptor agonism safely benefit diabetic patients, reducing the insulin requirements and glycemic excursions. Pramlintide is the triple proline human amylin analogue first used as injectable drug, but lacking physico-chemical compatibility when co-formulated with insulin. Here, we report the design and characterization of polymeric microparticles for oral delivery of pramlintide. Eudragit S100, a gastric-resistant polymer, was used in preparation of pramlintide-loaded spherical microcapsules by double emulsion and solvent evaporation technique, with approximately 66 μm ± 11 particle size, with 83.2% ± 2.7 efficiency for pramlintide entrapment and 67.6% ± 2.1 yield. Intra-venous pramlintide free in solution showed a plasmatic half-life of 6.8 min in mice. In contrast, oral delivery of acid-resistant pramlintide-loaded microparticles in mice showed a protracted release for 120 min compared to 30 min obtained for pramlintide in solution. Our data provide evidences for the potential use of the oral route in the therapeutic development of pramlintide formulations.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"50 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139656258","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A well-established mechanism of action for managing pneumonia using Glycyrrhiza is unknown. Using network pharmacology and molecular docking simulations, we investigated the mechanism of action of Glycyrrhiza against pneumonia. To identify the targets of the active components of Glycyrrhiza from the Traditional Chinese Medicine Systems Pharmacology database, oral bioavailability and drug likeness were utilized as indicators. Pneumonia-associated genes were identified and screened from the databases. Integrated analysis was conducted to elucidate the relationship between the active components of Glycyrrhiza and intersecting genes; a comprehensive Glycyrrhiza active component-target gene relationship map was constructed. Intersecting genes underwent Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses to examine their biological functions. A protein–protein interaction network map was constructed to identify hub genes. Molecular docking simulations were performed to investigate binding interactions between hub genes and their corresponding active components. Of the 96 overlapping genes, topological analysis revealed 10 hub genes. Glycyrrhiza exerts therapeutic effects through a multi-target and multipathway approach, suggesting a synergistic treatment for pneumonia. MAPK14 showed a favorable binding affinity with most of the active compounds, indicating that MAPK14 and related compounds in Glycyrrhiza have development potential.
{"title":"Network pharmacology combined with molecular docking simulations reveal the mechanism of action of Glycyrrhiza for treating pneumonia","authors":"Dongxin Yang, Zhehong Li, Yongrui Peng, Xiaofeng Zhu, Jun Gong, Cuilian Chen","doi":"10.1002/pep2.24342","DOIUrl":"https://doi.org/10.1002/pep2.24342","url":null,"abstract":"A well-established mechanism of action for managing pneumonia using <i>Glycyrrhiza</i> is unknown. Using network pharmacology and molecular docking simulations, we investigated the mechanism of action of <i>Glycyrrhiza</i> against pneumonia. To identify the targets of the active components of <i>Glycyrrhiza</i> from the Traditional Chinese Medicine Systems Pharmacology database, oral bioavailability and drug likeness were utilized as indicators. Pneumonia-associated genes were identified and screened from the databases. Integrated analysis was conducted to elucidate the relationship between the active components of <i>Glycyrrhiza</i> and intersecting genes; a comprehensive <i>Glycyrrhiza</i> active component-target gene relationship map was constructed. Intersecting genes underwent Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analyses to examine their biological functions. A protein–protein interaction network map was constructed to identify hub genes. Molecular docking simulations were performed to investigate binding interactions between hub genes and their corresponding active components. Of the 96 overlapping genes, topological analysis revealed 10 hub genes. <i>Glycyrrhiza</i> exerts therapeutic effects through a multi-target and multipathway approach, suggesting a synergistic treatment for pneumonia. MAPK14 showed a favorable binding affinity with most of the active compounds, indicating that MAPK14 and related compounds in <i>Glycyrrhiza</i> have development potential.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"47 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139496065","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cong Shibo, Yu Tong, Wang Sili, Gu Shuxiao, Qiao Yanfang, Liu Susu, Gao Pan, Meng Jingke, Chai Xinlou
This study aims to explore the molecular mechanism and potential active ingredients of Danhe granules in improving the blood lipid level of hypercholesterolemia by intestinal TICE (transintestinal cholesterol excretion) through in vivo experimental research, network pharmacology methods, and molecular docking. The diet-induced hypercholesterolemia rat model evaluated the effects of Danhe granules on fecal cholesterol levels and intestinal TICE-related protein expression in rats. Based on the experimental results, the network pharmacology method was used to predict the potential active ingredients, and the binding strength between the potential active ingredients and key targets was further confirmed by molecular docking. Danhe granules increased LDLR, ABCG5, ABCB1, and LXRα which can promote the uptake of cholesterol by intestinal cells and the excretion of cholesterol into the intestine, and then reduce the levels of serum LDL-C and TC and increase the level of fecal TC, so as to achieve the effect of treating hypercholesterolemia. The results of network pharmacology and molecular docking showed that 19 active ingredients had good binding activity with the targets. These may be the active ingredients of Danhe granules for the treatment of hypercholesterolemia. This study revealed the molecular mechanism of Danhe granules in the treatment of hypercholesterolemia via TICE, and preliminarily clarified the potential effective ingredients. It provides new ideas for the treatment of hypercholesterolemia and the development of new drugs.
{"title":"Molecular mechanism study and potential active ingredients prediction of Danhe granules in the treatment of hypercholesterolemia via TICE","authors":"Cong Shibo, Yu Tong, Wang Sili, Gu Shuxiao, Qiao Yanfang, Liu Susu, Gao Pan, Meng Jingke, Chai Xinlou","doi":"10.1002/pep2.24345","DOIUrl":"https://doi.org/10.1002/pep2.24345","url":null,"abstract":"This study aims to explore the molecular mechanism and potential active ingredients of Danhe granules in improving the blood lipid level of hypercholesterolemia by intestinal TICE (transintestinal cholesterol excretion) through in vivo experimental research, network pharmacology methods, and molecular docking. The diet-induced hypercholesterolemia rat model evaluated the effects of Danhe granules on fecal cholesterol levels and intestinal TICE-related protein expression in rats. Based on the experimental results, the network pharmacology method was used to predict the potential active ingredients, and the binding strength between the potential active ingredients and key targets was further confirmed by molecular docking. Danhe granules increased LDLR, ABCG5, ABCB1, and LXRα which can promote the uptake of cholesterol by intestinal cells and the excretion of cholesterol into the intestine, and then reduce the levels of serum LDL-C and TC and increase the level of fecal TC, so as to achieve the effect of treating hypercholesterolemia. The results of network pharmacology and molecular docking showed that 19 active ingredients had good binding activity with the targets. These may be the active ingredients of Danhe granules for the treatment of hypercholesterolemia. This study revealed the molecular mechanism of Danhe granules in the treatment of hypercholesterolemia via TICE, and preliminarily clarified the potential effective ingredients. It provides new ideas for the treatment of hypercholesterolemia and the development of new drugs.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"51 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139459578","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Michael A. Lee, Joseph S. Brown, Andrei Loas, Bradley L. Pentelute
Solid-phase peptide synthesis (SPPS) is widely used to produce peptides. Since its invention, the solid support has enabled the elongation of the synthetic peptide chain. As technologies have evolved, the length of peptide chains accessible with SPPS has grown to that of single-domain proteins. Resins for SPPS have advanced to improve synthesis capabilities as well. The functionalization of solid supports with polyethylene glycol (PEG) is commonly employed in a range of commercially available resins to aid in the ability to synthesize long or difficult sequences. A notable example of a widely used PEG-based solid support is ChemMatrix® resin; however, this and several similar resins have recently been discontinued. Here, we demonstrate and compare the capabilities of OctaGel™, ProTide®, and TentaGel XV® resins in synthesizing sequences ranging from peptides to single domain proteins using automated fast-flow peptide synthesis. Our studies indicate that each resin performs well for routine peptide synthesis by automated flow, whereas TentaGel XV resin showed the best performance for synthesis of difficult or long peptide sequences when comparing quality using yield, purity, and real-time UV absorbance monitoring during synthesis.
{"title":"Investigation of commercially available resins for the automated flow synthesis of difficult or long peptide sequences","authors":"Michael A. Lee, Joseph S. Brown, Andrei Loas, Bradley L. Pentelute","doi":"10.1002/pep2.24344","DOIUrl":"https://doi.org/10.1002/pep2.24344","url":null,"abstract":"Solid-phase peptide synthesis (SPPS) is widely used to produce peptides. Since its invention, the solid support has enabled the elongation of the synthetic peptide chain. As technologies have evolved, the length of peptide chains accessible with SPPS has grown to that of single-domain proteins. Resins for SPPS have advanced to improve synthesis capabilities as well. The functionalization of solid supports with polyethylene glycol (PEG) is commonly employed in a range of commercially available resins to aid in the ability to synthesize long or difficult sequences. A notable example of a widely used PEG-based solid support is ChemMatrix® resin; however, this and several similar resins have recently been discontinued. Here, we demonstrate and compare the capabilities of OctaGel™, ProTide®, and TentaGel XV® resins in synthesizing sequences ranging from peptides to single domain proteins using automated fast-flow peptide synthesis. Our studies indicate that each resin performs well for routine peptide synthesis by automated flow, whereas TentaGel XV resin showed the best performance for synthesis of difficult or long peptide sequences when comparing quality using yield, purity, and real-time UV absorbance monitoring during synthesis.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"87 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139459828","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Qiongqiong Ren, Zejun Fan, Rong Han, Meihui Sang, Changxing Ma, Xiaoli Zhao, Shenlin Wang
Engineered Escherichia coli (E. coli) strains have been widely used to produce isotopically labeled peptides for NMR characterization on their structures and interactions. However, production of antimicrobial peptides (AMPs) by E. coli is still challenging, because AMPs are toxic to E. coli host and would lead to cell death after induction. On the other hand, expression of short peptides in E. coli host often encounter problems of the short in vivo lifetime of the peptides, which were rapidly degraded by endogenous enzymes during expression and purification steps. This report presents a practical method for overcoming these bottlenecks to enable E. coli to express AMPs and peptides that have short in vivo lifetime. This design uses the fusion of thioredoxin tags at both the N- and C-termini of the target peptides. The steric effect of the large soluble tags at both ends of the peptide reduces peptide accessibility, thereby enhancing their in vivo stability and eliminating the toxicity associated with AMPs. The approach was validated using an AMP A3K/L7K-LAH4 (K3K7) and a membrane fusion peptide (FP), which is a segment of the spike protein of SARS-CoV-2 and functions in fusing viral membranes and host cell membranes. Fusion expression of K3K7 with a thioredoxin tag only at the N-terminal resulted in high toxicity to the host cells, leading to impaired cell growth and a failure to obtain expressed fusion protein. In contrast, the fusion proteins from both termini were successfully expressed and purified. In the case of the FP, the fusion of thioredoxin at both termini significantly enhanced its stability, protecting it from enzymatic degradation during expression and purification steps. On the contrary, the FP with thioredoxin fused only at the N-terminal was found to be unstable in E. coli host strains. As stable isotope labeling on peptide is essentially important in NMR-based structure and interaction studies, we also demonstrated that the developed approach enables efficient 15N labeling for NMR studies. This strategy may also be extended to produce other challenging peptides.
工程大肠杆菌（E. coli）菌株已被广泛用于生产同位素标记的肽，以对其结构和相互作用进行核磁共振表征。然而，用大肠杆菌生产抗菌肽（AMPs）仍然具有挑战性，因为 AMPs 对大肠杆菌宿主有毒，诱导后会导致细胞死亡。另一方面，在大肠杆菌宿主中表达短肽往往会遇到短肽在体内寿命短的问题，因为短肽在表达和纯化过程中会被内源酶迅速降解。本报告提出了一种克服这些瓶颈的实用方法，使大肠杆菌能够表达体内寿命短的 AMP 和多肽。这种设计在目标肽的 N 端和 C 端融合了硫代毒素标签。肽两端的大型可溶性标签的立体效应降低了肽的可及性，从而提高了肽在体内的稳定性，并消除了与 AMP 相关的毒性。该方法使用 AMP A3K/L7K-LAH4 (K3K7) 和膜融合肽 (FP) 进行了验证，膜融合肽是 SARS-CoV-2 的尖峰蛋白的一个片段，具有融合病毒膜和宿主细胞膜的功能。K3K7 的融合表达仅在 N 端带有硫代毒素标签，因此对宿主细胞的毒性很高，导致细胞生长受阻，无法获得表达的融合蛋白。相比之下，两个末端的融合蛋白都能成功表达和纯化。就 FP 而言，硫氧还蛋白在两个末端的融合大大提高了其稳定性，使其在表达和纯化步骤中不会被酶降解。相反，只在 N 端融合了硫氧还蛋白的 FP 在大肠杆菌宿主菌株中不稳定。在基于核磁共振的结构和相互作用研究中，肽上稳定的同位素标记非常重要，因此我们也证明了所开发的方法能够为核磁共振研究提供高效的 15N 标记。这种策略也可扩展到其他具有挑战性的多肽的制备中。
{"title":"A strategy for producing isotopically labeled peptides with antimicrobial activity or with short in vivo lifetime in Escherichia coli","authors":"Qiongqiong Ren, Zejun Fan, Rong Han, Meihui Sang, Changxing Ma, Xiaoli Zhao, Shenlin Wang","doi":"10.1002/pep2.24340","DOIUrl":"https://doi.org/10.1002/pep2.24340","url":null,"abstract":"Engineered <i>Escherichia coli</i> (<i>E. coli</i>) strains have been widely used to produce isotopically labeled peptides for NMR characterization on their structures and interactions. However, production of antimicrobial peptides (AMPs) by <i>E. coli</i> is still challenging, because AMPs are toxic to <i>E. coli</i> host and would lead to cell death after induction. On the other hand, expression of short peptides in <i>E. coli</i> host often encounter problems of the short in vivo lifetime of the peptides, which were rapidly degraded by endogenous enzymes during expression and purification steps. This report presents a practical method for overcoming these bottlenecks to enable <i>E. coli</i> to express AMPs and peptides that have short in vivo lifetime. This design uses the fusion of thioredoxin tags at both the N- and C-termini of the target peptides. The steric effect of the large soluble tags at both ends of the peptide reduces peptide accessibility, thereby enhancing their in vivo stability and eliminating the toxicity associated with AMPs. The approach was validated using an AMP A3K/L7K-LAH4 (K3K7) and a membrane fusion peptide (FP), which is a segment of the spike protein of SARS-CoV-2 and functions in fusing viral membranes and host cell membranes. Fusion expression of K3K7 with a thioredoxin tag only at the N-terminal resulted in high toxicity to the host cells, leading to impaired cell growth and a failure to obtain expressed fusion protein. In contrast, the fusion proteins from both termini were successfully expressed and purified. In the case of the FP, the fusion of thioredoxin at both termini significantly enhanced its stability, protecting it from enzymatic degradation during expression and purification steps. On the contrary, the FP with thioredoxin fused only at the N-terminal was found to be unstable in <i>E. coli</i> host strains. As stable isotope labeling on peptide is essentially important in NMR-based structure and interaction studies, we also demonstrated that the developed approach enables efficient <sup>15</sup>N labeling for NMR studies. This strategy may also be extended to produce other challenging peptides.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"16 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139459579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The coronavirus 2 (SARS‐CoV‐2), a positive‐sense RNA virus that causes severe acute respiratory illness, is the cause of Coronavirus Disease 2019 (COVID‐19) and Monkeypox is a zoonotic viral disease caused by a double‐strand enveloped DNA virus, a member of the Poxviridae family under the umbrella of the Orthopoxvirus genus. These viruses have harmed global health, societal stability, and global economy prompting the development of new therapeutic options. The purpose of this study was to analyze the potency of hazelnut‐derived bioactive peptides for usage in preventive measures against SARS‐CoV‐2 and Monkeypox infections. Since SARS‐CoV‐2 entry into the host cell takes place through angiotensin‐converting enzyme (ACE) receptors, an in silico attempt was made here to analyze the previously characterized ACE‐inhibitory hazelnut peptides for their anti‐COVID potential. First of all, peptide sequences with PeptideRanker values >0.5 were determined, where 43 of 256 hazelnut peptides met this condition. BIOPEP tools were used to calculate their ACE‐inhibitory characteristics. VPHW and DENPRHF demonstrated the strongest ACE‐inhibitory activity based on BIOPEP analysis. The binding potentials of VPHW (p < 0.001) and DENPRHF (p < 0.001) peptides to ACE were statistically significant based on PepSite2 analysis. The hazelnut‐derived peptides were docked with ACE, spike proteins, SARS‐CoV‐2 proteases, host cell receptors, and Monkeypox 2022 membrane proteins using HPEPDOCK and CABS‐dock. The in silico findings pointed out potential inhibition of SARS‐CoV‐2 main protease in the host cell and Monkeypox 2022 membrane protein and demonstration of multiple bioactivities.
{"title":"In silico prediction of preventive activities of bioactive hazelnut peptides against COVID‐19 and Monkeypox 2022","authors":"Zeynep Saliha Güneş, B. Cakir, İbrahim Gülseren","doi":"10.1002/pep2.24341","DOIUrl":"https://doi.org/10.1002/pep2.24341","url":null,"abstract":"The coronavirus 2 (SARS‐CoV‐2), a positive‐sense RNA virus that causes severe acute respiratory illness, is the cause of Coronavirus Disease 2019 (COVID‐19) and Monkeypox is a zoonotic viral disease caused by a double‐strand enveloped DNA virus, a member of the Poxviridae family under the umbrella of the Orthopoxvirus genus. These viruses have harmed global health, societal stability, and global economy prompting the development of new therapeutic options. The purpose of this study was to analyze the potency of hazelnut‐derived bioactive peptides for usage in preventive measures against SARS‐CoV‐2 and Monkeypox infections. Since SARS‐CoV‐2 entry into the host cell takes place through angiotensin‐converting enzyme (ACE) receptors, an in silico attempt was made here to analyze the previously characterized ACE‐inhibitory hazelnut peptides for their anti‐COVID potential. First of all, peptide sequences with PeptideRanker values >0.5 were determined, where 43 of 256 hazelnut peptides met this condition. BIOPEP tools were used to calculate their ACE‐inhibitory characteristics. VPHW and DENPRHF demonstrated the strongest ACE‐inhibitory activity based on BIOPEP analysis. The binding potentials of VPHW (p < 0.001) and DENPRHF (p < 0.001) peptides to ACE were statistically significant based on PepSite2 analysis. The hazelnut‐derived peptides were docked with ACE, spike proteins, SARS‐CoV‐2 proteases, host cell receptors, and Monkeypox 2022 membrane proteins using HPEPDOCK and CABS‐dock. The in silico findings pointed out potential inhibition of SARS‐CoV‐2 main protease in the host cell and Monkeypox 2022 membrane protein and demonstration of multiple bioactivities.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"1 2","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139439501","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Shreya Banerjee, Sunil K. Gupta, Sunit Pal, Erode N. Prabhakaran
CisPro/transPro isomerism at the prolyl amide bond is a fundamental dynamism governing protein folding, structure, and functions. Since cisPro crystal structures are rare, the interactions influencing their structures are less understood, unlike transPro. Crystal data for the sterically hindered pivaloyl-cisProlyl amide bond (2,2-dimethyl-1-(1-pyrrolidinyl)-1-propanone, Piv-cisPro) were particularly lacking for decades. Here we introduce Piv-Pro-Xaa-OMe dipeptides which crystallize with the elusive Piv-cisPro (Xaa is Leu/Ile) and the abundant Piv-transPro (Xaa is Gly/Phe) conformers.
{"title":"Crystal structures reveal that the sterically hindered pivaloyl-cisProlyl amide bond is energetically frustrated","authors":"Shreya Banerjee, Sunil K. Gupta, Sunit Pal, Erode N. Prabhakaran","doi":"10.1002/pep2.24337","DOIUrl":"https://doi.org/10.1002/pep2.24337","url":null,"abstract":"CisPro/transPro isomerism at the prolyl amide bond is a fundamental dynamism governing protein folding, structure, and functions. Since cisPro crystal structures are rare, the interactions influencing their structures are less understood, unlike transPro. Crystal data for the sterically hindered pivaloyl-cisProlyl amide bond (2,2-dimethyl-1-(1-pyrrolidinyl)-1-propanone, Piv-cisPro) were particularly lacking for decades. Here we introduce Piv-Pro-Xaa-OMe dipeptides which crystallize with the elusive Piv-cisPro (Xaa is Leu/Ile) and the abundant Piv-transPro (Xaa is Gly/Phe) conformers.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"3 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139398069","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Seul Ki Yun, Seung Mok Yang, Moon Hwa Kwak, Jae Myung Park
Cancer-targeting diagnostics should have a sensitive and specific binding affinity. To achieve this, biomarker development is critical. This study aimed to develop and validate a 7-mer cyclic peptide probe that can target gastric cancer. We developed this probe based on LGR5 (leucine-rich repeat-containing G-protein coupled receptor 5)-specific targeting, which is a marker for gastric cancer stem cells. An LGR5 targeting peptide sequence that was developed using phage display technology resulted in a cyclic peptide, C-YLASRVH-C (named YLA). We conjugated this peptide with fluorescent probes to validate its specific targeting ability for gastric cancer. The fluorescence-labeled YLA peptide exhibited 3.0-fold higher fluorescence intensity in a gastric cancer cell line (MKN45) than it did in a normal cell line (CCD841 cells). In contrast, pancreatic and colorectal cancer cells did not show significant fluorescence intensity with the YLA peptide. To verify its tumor-targeting affinity, we developed a control peptide, C-YLASAVH-C (named YLASA) using an ALA scanning experiment. Whole-body imaging of a gastric cancer xenograft model showed higher fluorescence intensity of tumors in the YLA peptide group than in the control peptide group. Moreover, ex vivo imaging of tumor tissues exhibited 6.8-fold higher fluorescence intensity in the YLA peptide group compared to that in the YLASA control peptide group. In conclusion, we confirmed that the YLA peptide probe functions as a specific diagnostic probe for gastric cancer. We anticipate that it will play a theranostic role through further development.
{"title":"Development and validation of cyclic peptide probe for gastric cancer based on phage display technique","authors":"Seul Ki Yun, Seung Mok Yang, Moon Hwa Kwak, Jae Myung Park","doi":"10.1002/pep2.24339","DOIUrl":"https://doi.org/10.1002/pep2.24339","url":null,"abstract":"Cancer-targeting diagnostics should have a sensitive and specific binding affinity. To achieve this, biomarker development is critical. This study aimed to develop and validate a 7-mer cyclic peptide probe that can target gastric cancer. We developed this probe based on LGR5 (leucine-rich repeat-containing G-protein coupled receptor 5)-specific targeting, which is a marker for gastric cancer stem cells. An LGR5 targeting peptide sequence that was developed using phage display technology resulted in a cyclic peptide, C-YLASRVH-C (named YLA). We conjugated this peptide with fluorescent probes to validate its specific targeting ability for gastric cancer. The fluorescence-labeled YLA peptide exhibited 3.0-fold higher fluorescence intensity in a gastric cancer cell line (MKN45) than it did in a normal cell line (CCD841 cells). In contrast, pancreatic and colorectal cancer cells did not show significant fluorescence intensity with the YLA peptide. To verify its tumor-targeting affinity, we developed a control peptide, C-YLASAVH-C (named YLASA) using an ALA scanning experiment. Whole-body imaging of a gastric cancer xenograft model showed higher fluorescence intensity of tumors in the YLA peptide group than in the control peptide group. Moreover, ex vivo imaging of tumor tissues exhibited 6.8-fold higher fluorescence intensity in the YLA peptide group compared to that in the YLASA control peptide group. In conclusion, we confirmed that the YLA peptide probe functions as a specific diagnostic probe for gastric cancer. We anticipate that it will play a theranostic role through further development.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"23 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139094506","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Gabriella Rodrigues Gonçalves, Marciele Souza Silva, Layrana Azevedo dos Santos, Larissa Maximiniano Resende, Gabriel Bonan Taveira, Thomas Zacarone Afonso Guimarães, Sarah Rodrigues Ferreira, Antonia Elenir Amancio Oliveira, Celso Shiniti Nagano, Renata Pinheiro Chaves, André de Oliveira Carvalho, Rosana Rodrigues, Olney Vieira da Motta, Valdirene Moreira Gomes
In recent years, there have been several reports of the presence of toxic proteins in cultivated or wild plant species, which are implicated in plant defense mechanisms. The existence of these proteins raises the possibility of biotechnological applications originating from the development of new techniques to combat diseases caused by fungi. In this context, there are chitin-binding proteins. Chitin is an essential component of the fungal cell wall, so chitin-binding proteins are important in controlling fungal growth. Thus, the objective of this study was to characterize and evaluate the in vitro antimicrobial effect of peptides with chitin binding properties isolated from Capsicum annuum seeds on the growth of the genus Candida. Initially, proteins were extracted in phosphate pH 5.4, and a chitin column was equilibrated with sodium acetate (0.08 M, pH 4.5), where 50 mg of the peptide-rich heated fraction from each species was applied. Subsequently, the retained material was eluted with 0.1 M HCl. Tricine SDS–PAGE was used to visualize the peptides. After chromatography, two fractions, F1 (not retained in the chitin column) and F2 (retained in the chitin column, named Ca-F2), were obtained. Electrophoresis showed major protein bands between 3 and 14 kDa. Electrophoresis from chitin affinity chromatography also showed major bands between 3 and 14 kDa, especially for Ca-F2 retained in the column. One peptide obtained from the F2 fraction was identified by mass spectrometry and showed similarity to seed 2S albumin, named Ca-Alb2S. Ca-F2 inhibited the growth of C. albicans and C. tropicalis, was not toxic to mammalian cells and still had a high survival rate when tested in vivo on Galleria mellonella larvae. This is the first report of chitin-binding peptides isolated from Capsicum seeds through an affinity column and their biological activities. These studies are at an early stage; therefore, other tests are needed to study the mechanism of action of the fraction, since the findings indicate great potential for the development of new antifungal molecules.
{"title":"Chitin-binding peptides from Capsicum annuum with antifungal activity and low toxicity to mammalian cells and Galleria mellonella larvae","authors":"Gabriella Rodrigues Gonçalves, Marciele Souza Silva, Layrana Azevedo dos Santos, Larissa Maximiniano Resende, Gabriel Bonan Taveira, Thomas Zacarone Afonso Guimarães, Sarah Rodrigues Ferreira, Antonia Elenir Amancio Oliveira, Celso Shiniti Nagano, Renata Pinheiro Chaves, André de Oliveira Carvalho, Rosana Rodrigues, Olney Vieira da Motta, Valdirene Moreira Gomes","doi":"10.1002/pep2.24338","DOIUrl":"https://doi.org/10.1002/pep2.24338","url":null,"abstract":"In recent years, there have been several reports of the presence of toxic proteins in cultivated or wild plant species, which are implicated in plant defense mechanisms. The existence of these proteins raises the possibility of biotechnological applications originating from the development of new techniques to combat diseases caused by fungi. In this context, there are chitin-binding proteins. Chitin is an essential component of the fungal cell wall, so chitin-binding proteins are important in controlling fungal growth. Thus, the objective of this study was to characterize and evaluate the <i>in vitro</i> antimicrobial effect of peptides with chitin binding properties isolated from <i>Capsicum annuum</i> seeds on the growth of the genus <i>Candida</i>. Initially, proteins were extracted in phosphate pH 5.4, and a chitin column was equilibrated with sodium acetate (0.08 M, pH 4.5), where 50 mg of the peptide-rich heated fraction from each species was applied. Subsequently, the retained material was eluted with 0.1 M HCl. Tricine SDS–PAGE was used to visualize the peptides. After chromatography, two fractions, F1 (not retained in the chitin column) and F2 (retained in the chitin column, named <i>Ca</i>-F2), were obtained. Electrophoresis showed major protein bands between 3 and 14 kDa. Electrophoresis from chitin affinity chromatography also showed major bands between 3 and 14 kDa, especially for <i>Ca</i>-F2 retained in the column. One peptide obtained from the F2 fraction was identified by mass spectrometry and showed similarity to seed 2S albumin, named <i>Ca</i>-Alb2S. <i>Ca</i>-F2 inhibited the growth of <i>C. albicans</i> and <i>C. tropicalis</i>, was not toxic to mammalian cells and still had a high survival rate when tested in vivo on <i>Galleria mellonella</i> larvae. This is the first report of chitin-binding peptides isolated from <i>Capsicum</i> seeds through an affinity column and their biological activities. These studies are at an early stage; therefore, other tests are needed to study the mechanism of action of the fraction, since the findings indicate great potential for the development of new antifungal molecules.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"131 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2024-01-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139094503","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Nicholas A. Dalley, Kimberlee L. Stern, Richard R. Kitchen, Keegan B. Lloyd, Joshua L. Price
Coiled coils are one of most common protein quaternary structures and represent the best understood relationship between amino acid sequence and protein conformation. Whereas the roles of residues at the canonical heptad positions the a, d, e, and g are understood in precise detail, conventional approaches often assume that the solvent-exposed b-, c-, and f-positions can be varied broadly for application-specific purposes with minimal consequences. However, a growing body of evidence suggests that interactions among these b, c, and f residues can contribute substantially to coiled-coil conformational stability. In the trimeric coiled coil described here, we find that b-position Glu10 engages in a stabilizing long-range synergistic interaction with c-position Lys18 (ΔΔΔGf = −0.65 ± 0.02 kcal/mol). This favorable interaction depends strongly on the presence of two nearby f-position residues: Lys 7 and Tyr14. Extensive mutational analysis of these residues in the presence of added salt versus denaturant suggests that this long-range synergistic interaction is primarily electrostatic in origin, but also depends on the precise location and acidity of a side-chain hydrogen-bond donor within f-position Tyr14.
{"title":"Electrostatic origin of a stabilizing synergistic interaction among b-, c-, and f-residues in a trimeric coiled coil","authors":"Nicholas A. Dalley, Kimberlee L. Stern, Richard R. Kitchen, Keegan B. Lloyd, Joshua L. Price","doi":"10.1002/pep2.24336","DOIUrl":"https://doi.org/10.1002/pep2.24336","url":null,"abstract":"Coiled coils are one of most common protein quaternary structures and represent the best understood relationship between amino acid sequence and protein conformation. Whereas the roles of residues at the canonical heptad positions the <i>a</i>, <i>d</i>, <i>e</i>, and <i>g</i> are understood in precise detail, conventional approaches often assume that the solvent-exposed <i>b</i>-, <i>c</i>-, and <i>f</i>-positions can be varied broadly for application-specific purposes with minimal consequences. However, a growing body of evidence suggests that interactions among these <i>b</i>, <i>c</i>, and <i>f</i> residues can contribute substantially to coiled-coil conformational stability. In the trimeric coiled coil described here, we find that <i>b</i>-position Glu10 engages in a stabilizing long-range synergistic interaction with <i>c</i>-position Lys18 (ΔΔΔ<i>G</i><sub><i>f</i></sub> = −0.65 ± 0.02 kcal/mol). This favorable interaction depends strongly on the presence of two nearby <i>f</i>-position residues: Lys 7 and Tyr14. Extensive mutational analysis of these residues in the presence of added salt versus denaturant suggests that this long-range synergistic interaction is primarily electrostatic in origin, but also depends on the precise location and acidity of a side-chain hydrogen-bond donor within <i>f</i>-position Tyr14.","PeriodicalId":19825,"journal":{"name":"Peptide Science","volume":"16 1","pages":""},"PeriodicalIF":2.4,"publicationDate":"2023-12-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"138518134","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
京公网安备 11010802042870号
京ICP备2023020795号-1
扫码关注我们
求助内容：
应助结果提醒方式：