Protein and Peptide Letters最新文献

Enhancing Tissue Factor Production: The Role of N-Glycosylation and ERAD Pathway Modulation. 促进组织因子的产生：n -糖基化和ERAD通路调节的作用。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-27 DOI: 10.2174/0109298665364078250519065417

Yi-Shi Liu, Yue Dou, Xiaoman Zhou, Zijie Li, Nakanishi Hideki

{"title":"Enhancing Tissue Factor Production: The Role of N-Glycosylation and ERAD Pathway Modulation.","authors":"Yi-Shi Liu, Yue Dou, Xiaoman Zhou, Zijie Li, Nakanishi Hideki","doi":"10.2174/0109298665364078250519065417","DOIUrl":"https://doi.org/10.2174/0109298665364078250519065417","url":null,"abstract":"Background: Tissue Factor (TF) is a crucial transmembrane glycoprotein that triggers blood coagulation upon vascular or tissue injury by binding to plasma factors VII and VIIa. In recent years, the demand for TF has rapidly increased due to its pivotal role in preoperative coagulation tests. However, large-scale production of TF remains challenging despite successful recombinant expression, as incorrect post-translational modifications adversely affect TF activity.Objective: This study aims to investigate the role of post-translational modifications, specifically N-glycosylation, in TF activity and stability. Additionally, it explores strategies to enhance TF production by reducing its degradation through genetic modulation.Methods: We compared TF activity derived from human cells and E. coli to assess the impact of post-translational modifications. Furthermore, we examined the effect of N-glycosylation on TF function. To address TF degradation, we knocked out the HRD1 gene, a key component of the endoplasmic- reticulum-associated degradation (ERAD) pathway, and evaluated its impact on TF stability and activity.Results: TF produced in human cells exhibited higher activity than TF expressed in E. coli, emphasizing the importance of post-translational modifications. Specifically, N-glycosylation was found to influence TF activity and stability significantly. Additionally, we observed that knocking out the HRD1 gene effectively reduced TF degradation without compromising its activity.Conclusion: Our findings underscore the crucial role of N-glycosylation in TF function and stability. Moreover, the modulation of the ERAD pathway through HRD1 knockout presents a promising approach for enhancing TF production. These insights could contribute to the large-scale manufacturing of functionally active TF for clinical and research applications.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144187769","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}

引用次数: 0

Molecular Cloning and Expression of Cryptocyanin Gene Isolated from an Indian Variety of Scylla olivacea. 印度橄榄Scylla品种隐花色素基因的克隆与表达。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-21 DOI: 10.2174/0109298665387863250506105601

Simran Mann, Chittibabu Shanthi, Manu Asthana

{"title":"Molecular Cloning and Expression of Cryptocyanin Gene Isolated from an Indian Variety of Scylla olivacea.","authors":"Simran Mann, Chittibabu Shanthi, Manu Asthana","doi":"10.2174/0109298665387863250506105601","DOIUrl":"https://doi.org/10.2174/0109298665387863250506105601","url":null,"abstract":"Background: Molting and reproduction play vital roles in the life cycle of brachyuran crabs, and these two processes are closely interconnected. A key player in the molting cycle is cryptocyanin, which is similar to hemocyanin in sequence, size, and structure. Hemocyanin is a copper-containing oxygen-binding protein, while cryptocyanin is a copper-free protein that lacks oxygen-binding capacity.Objective: The goal of the study was to carry out the isolation, cloning, and expression of the partial cryptocyanin gene from the Indian variety of Scylla olivacea.Methods: The partial cryptocyanin gene was isolated from the hemocytes of the S. olivacea male and female crabs by qPCR for comparative expression analysis of the cryptocyanin gene.Results: We successfully amplified, cloned, and expressed a 519bp partial cDNA encoding cryptocyanin from the Indian variety of Scylla olivacea, within the pRSET-B vector.Conclusion: Our study showed that crustacean cDNA can be effectively expressed in bacterial vectors, and clones were stable for up to 6 months at -80oC. Real-time data showed a significant difference in cryptocyanin levels between male and female crabs. This finding highlights the need for further research with a larger sample size for better understanding.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144143106","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}

引用次数: 0

Design, Expression, and Purification of a Fusion Enzyme Containing Terminal Deoxynucleotidyl Transferase from B. bovis and DNA-Binding Proteins from E. coli. 含有牛B.末端脱氧核苷酸转移酶和大肠杆菌dna结合蛋白的融合酶的设计、表达和纯化。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-21 DOI: 10.2174/0109298665372636250504084653

Antos Sachanka, Veronika Shchur, Yaraslau Dzichenka, Aleksei Yantsevich

{"title":"Design, Expression, and Purification of a Fusion Enzyme Containing Terminal Deoxynucleotidyl Transferase from B. bovis and DNA-Binding Proteins from E. coli.","authors":"Antos Sachanka, Veronika Shchur, Yaraslau Dzichenka, Aleksei Yantsevich","doi":"10.2174/0109298665372636250504084653","DOIUrl":"https://doi.org/10.2174/0109298665372636250504084653","url":null,"abstract":"Background: Gene fusion techniques have yielded promising results in the fusion of thermostable polymerases (Taq and Pfu) with single-stranded and double-stranded DNA-binding proteins. Constructing a terminal deoxynucleotidyl transferase (TdT) fusion enzyme with DNAbinding protein domains can enhance thermostability and broaden the enzyme's application field. This makes it a promising candidate for cost-effective de novo DNA synthesis and a more effective tool for demonstrating apoptosis and detecting viral DNA/RNA.Methods: The design of fusion proteins was based on molecular dynamics and homology modeling. Native and fusion proteins were isolated using affinity chromatography on HisTrap HP. Thermostability was assessed through differential scanning fluorimetry and dynamic light scattering. HPLC analysis was conducted to evaluate enzyme activity.Results: According to the in silico predictions of the fusion protein structure, a homotetramer was formed. The expressed fusion proteins were successfully purified under native conditions, similar to TdT. The total yields of the studied proteins were 130 mg/L for single-stranded binding protein from E. coli (EcSSB), 5 mg/L for TdT, 9 mg/L for TdT_L1_EcSSB, and 7 mg/L for TdT_L2_EcSSB. The measured radius of TdT (3.5 nm) was found to be consistent with a monomeric structure; however, the fusion proteins were expected to form a homotetramer. Additionally, fusion with EcSSB was found to prevent aggregation, which positively affected the thermal stability of the fusion protein. Instead of elongating the substrate by adding nucleotides, the fusion enzyme removed a nucleotide, specifically TTP, from the 3'-end of the DNA strand.Conclusion: The fusion of TdT with EcSSB resulted in increased thermal stability and a reduced ability to add nucleotides to the substrate.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144142991","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}

引用次数: 0

Cell-Free Expression of HPV16 Minor Capsid Protein L2 and Its Interaction with S100A10. HPV16次要衣壳蛋白L2的细胞外表达及其与S100A10的相互作用。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-21 DOI: 10.2174/0109298665390494250513110604

Wenqi Jiang, Lian Wu, Xiangchun Shen, Qingshan Bill Fu

{"title":"Cell-Free Expression of HPV16 Minor Capsid Protein L2 and Its Interaction with S100A10.","authors":"Wenqi Jiang, Lian Wu, Xiangchun Shen, Qingshan Bill Fu","doi":"10.2174/0109298665390494250513110604","DOIUrl":"https://doi.org/10.2174/0109298665390494250513110604","url":null,"abstract":"Background: Human papillomavirus type 16 (HPV16) is implicated in various malignancies. The virus enters host cells through endocytosis, during which the minor capsid protein L2 interacts with the S100A10 subunit of the annexin A2 heterotetramer (A2t) on the host cell membrane. This interaction is critical for facilitating HPV entry and subsequent infection of human cells. Therefore, examining the interaction between the L2 protein and S100A10 is crucial for advancing our understanding of the mechanisms by which HPV16 infiltrates cells.Objective: The cell-free expression (CFE) system was investigated for L2 purification. The structure of L2 was characterized and its interaction with S100A10 was explored.Methods: The L2 protein was expressed using a CFE expression system, and its expression was verified via Western blotting. L2 was further purified through size-exclusion chromatography (SEC), and its structural features were preliminarily assessed using transmission electron microscopy (TEM) and circular dichroism (CD) spectroscopy. Additionally, surface plasmon resonance (SPR) was employed to analyze the interaction between L2 and S100A10.Results: Western blotting confirmed the successful expression of L2. TEM and CD provided preliminary structural observations of L2, and SPR measurements yielded precise kinetic parameters for the interaction between L2 and S100A10.Conclusion: In this study, we successfully expressed the HPV16 L2 protein using a cell-free protein expression system. Preliminary structural analysis using TEM and CD revealed key structural features of L2. Furthermore, SPR analysis provided detailed kinetic parameters for its interaction with S100A10. These findings provide more details on understanding L2's structural features, with broader implications for antipathogen studies.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144142985","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}

引用次数: 0

Virtual Screening of Phytoconstituents in Indian Spices Based on their Inhibitory Potential against SARS-CoV-2. 基于抑制SARS-CoV-2潜力的印度香料植物成分虚拟筛选

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-08 DOI: 10.2174/0109298665366911250416113831

Vaishali Singh, Aliza Rabbani, Veda P Pandey

{"title":"Virtual Screening of Phytoconstituents in Indian Spices Based on their Inhibitory Potential against SARS-CoV-2.","authors":"Vaishali Singh, Aliza Rabbani, Veda P Pandey","doi":"10.2174/0109298665366911250416113831","DOIUrl":"https://doi.org/10.2174/0109298665366911250416113831","url":null,"abstract":"Background: COVID-19 is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), a highly pathogenic human coronavirus (CoV). For the treatment of COVID-19, various drugs, ayurvedic formulations, used for other diseases, were repurposed. Ayurveda and yoga exhibited a pivotal role in the treatment of COVID-19. Various medicinal plants, including garlic, tulsi, clove, cinnamon, ginger, black pepper, and turmeric, are recommended for the prevention of COVID-19 as immunity boosters along with their antiviral property.Objective: In view of the drug repurposing approach, the present work has been initiated with the broader objectives of screening and identification of phytoconstituents of Indian spices against targets, namely furin, 3C-like protease (3CL-PRO), NSP-9 RNA binding protein, papain-like protease, RNA dependent RNA polymerase (RDRP), spike protein concerned with life cycle of SARS-CoV-2 using in-silico tools.Method: The phytoconstituents of Indian spices were screened for interaction with several targets using a molecular docking approach with the help of Discovery Studio 4.5 software. Furthermore, the pharmacokinetic analyses of selected ligands using ADMET and Lipinski's rule of five were also performed.Result: In the present study, more than 35 active phytoconstituents of Indian spices were screened for interaction with several identified targets of Covid-19 using a molecular docking approach. The ligands, namely morin, gingerol, myristic acid, quercetin, gallic acid, octacosanal, and alliin were found to be the top interacting ligands with the targets analyzed.Conclusion: Based on the present in-silico finding, the active components of spices could be considered for drug-lead compounds against COVID-19.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-08","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144008456","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}

引用次数: 0

Inhibition of Canonical Wnt Signaling in Renal Cell Carcinoma Bone Metastasis: An Immunohistochemical Analysis of DKK1 and LRP5 Expression. 典型Wnt信号在肾癌骨转移中的抑制作用：DKK1和LRP5表达的免疫组织化学分析。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-07 DOI: 10.2174/0109298665357331250416081850

Zixiong Huang, Yiqing Du, Huaqi Yin, Gongwei Wang, Tao Xu

{"title":"Inhibition of Canonical Wnt Signaling in Renal Cell Carcinoma Bone Metastasis: An Immunohistochemical Analysis of DKK1 and LRP5 Expression.","authors":"Zixiong Huang, Yiqing Du, Huaqi Yin, Gongwei Wang, Tao Xu","doi":"10.2174/0109298665357331250416081850","DOIUrl":"https://doi.org/10.2174/0109298665357331250416081850","url":null,"abstract":"Background and objective: Canonical Wnt (Wnt/β-catenin) signaling maintains bone homeostasis by promoting osteoblastic activities. The inhibitory factor, Dickkopf-1 (DKK1), enhances bone resorption in malignant diseases. Low-density lipoprotein-related protein (LRP) 5 is antagonized by DKK1. This study aimed to investigate the expression of DKK1 and LRP5 in renal cell carcinoma (RCC-BM).Methods: RCC-BM patients with paired samples of primary and metastatic lesions were selected for the study (RCC-BM group). RCC patients without any metastasis served as the control group (RCC-only group). Immunohistochemical staining with monoclonal anti-DKK1 and polyclonal anti- LRP5 antibody was conducted on paraffin-embedded slides. The staining results were recorded using scoring according to staining intensity in the renal tissue adjacent to the tumor, primary RCC lesions, and RCC-BM lesions.Results: DKK1 was differently expressed among normal renal tissues, primary RCC, and RCC- BM tissues (p<0.001). The DKK1 expression in primary RCC was significantly lower than that in normal renal tissues (p<0.001) without a difference between the RCC-BM and RCC-only groups. DKK1 expression in bone metastasis was significantly higher than that in primary tumors (p<0.001). For RCC-BM patients, the expression of LRP5 in the primary tumor was significantly lower than that in adjacent renal tissues (p<0.01). The tendency of lower expression was found in primary RCC from RCC-BM patients compared to RCC without metastasis (p=0.073).Conclusion: A \"rebound\" pattern of DKK1 expression in bone metastasis lesions and the decreasing LRP5 expression in primary lesions of RCC-BM patients suggested that Wnt/β-catenin signaling was inhibited in RCC-BM. The overexpression of DKK1 and reduced expression of LRP5 suggest that these markers may be useful for the early prediction of RCC-BM.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144028841","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}

引用次数: 0

Unraveling the Role of Functional Amyloids and Amyloid Peptides in Disease Detection. 揭示功能性淀粉样蛋白和淀粉样肽在疾病检测中的作用。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-05-07 DOI: 10.2174/0109298665368109250419175111

Priyanka Kumar, Nandini Sarkar

{"title":"Unraveling the Role of Functional Amyloids and Amyloid Peptides in Disease Detection.","authors":"Priyanka Kumar, Nandini Sarkar","doi":"10.2174/0109298665368109250419175111","DOIUrl":"https://doi.org/10.2174/0109298665368109250419175111","url":null,"abstract":"Amyloid refers to a specific quaternary structure characterized by fibrillar arrangements of proteins or peptides forming cross β-sheet architectures. Initially associated with diseases like Alzheimer's, amyloid was seen predominantly as pathological. However, recent research has revealed that amyloid also plays functional roles across various biological systems, from bacteria to mammals. The cross β-sheet structure of amyloid enables the transformation of soluble proteins into insoluble fibrils, providing high stability and a robust prion-like copying mechanism. However, recent research has revealed that amyloid also plays functional roles in various biological systems, such as biofilm formation in bacteria, aiding melanin biosynthesis in humans, and supporting the formation of fungal hyphae. Understanding the dual nature of amyloid-a pathological and functional entity-offers insights into disease mechanisms and therapeutic strategies. Recognizing the distinction between pathological and functional amyloids is crucial for advancing diagnostics and treatments. This review highlights the importance of functional amyloids (FAs), particularly in disease detection, underscoring their significant biological roles and potential applications.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-05-07","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144043891","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}

引用次数: 0

Development of a Peptide-Based Multiepitope Vaccine from the SARS-CoV-2 Spike Protein for Targeted Immune Response Against COVID-19. 基于SARS-CoV-2刺突蛋白多肽的靶向免疫应答疫苗的研制

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-04-14 DOI: 10.2174/0109298665364226250328084245

Thales Alves Campelo, Pedro Filho Noronha Souza, Daiane Maria Silva Brito, Cristiane Cunha Frota, Paulo Renato Zuquim Antas

{"title":"Development of a Peptide-Based Multiepitope Vaccine from the SARS-CoV-2 Spike Protein for Targeted Immune Response Against COVID-19.","authors":"Thales Alves Campelo, Pedro Filho Noronha Souza, Daiane Maria Silva Brito, Cristiane Cunha Frota, Paulo Renato Zuquim Antas","doi":"10.2174/0109298665364226250328084245","DOIUrl":"https://doi.org/10.2174/0109298665364226250328084245","url":null,"abstract":"Background: Since the Coronavirus Disease (COVID-19) became a pandemic in late 2019, vaccination remains the primary approach to combating the virus. Nevertheless, the emergence of new variants poses challenges to vaccine efficacy. This study aimed to identify targets within the SARS-CoV-2 spike (S) protein to detect T-cell responses to the five variants of concern from SARS-CoV-2: Alpha, Beta, Delta, Gamma, and Omicron.Methods: Here was employed immunoinformatics tools to develop a peptide-based vaccine targeting the spike protein of SARS-CoV-2 and its major variants, including Alpha, Beta, Delta, Gamma, and Omicron. The peptides were screened for antigenicity, toxicity, allergenicity, and physicochemical properties to ensure their safety and efficacy.Results: The potential T-cell epitopes with high immunogenicity and IFN-γ induction, are essential for a robust immune response by a comprehensive computational analysis. Population coverage analysis revealed significant coverage across diverse geographical regions, with significant efficacy in areas heavily impacted by the pandemic. Molecular docking simulations revealed strong interactions between the selected peptides and major histocompatibility complex class I (MHC-I) molecules, indicating their potential as vaccine candidates.Conclusion: Our study provides a systematic approach to the rational design of a peptide-based vaccine against COVID-19, providing insights for further experimental validation and development of effective vaccines.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-04-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144042507","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}

引用次数: 0

Use of Plant Peptidases for the Production of Therapeutic Peptides. 利用植物肽酶生产治疗性肽。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-04-11 DOI: 10.2174/0109298665373399250319082357

Cleverson Diniz Teixeira de Freitas, Jefferson Soares de Oliveira

{"title":"Use of Plant Peptidases for the Production of Therapeutic Peptides.","authors":"Cleverson Diniz Teixeira de Freitas, Jefferson Soares de Oliveira","doi":"10.2174/0109298665373399250319082357","DOIUrl":"https://doi.org/10.2174/0109298665373399250319082357","url":null,"abstract":"Peptidases play crucial roles in numerous physiological processes within living organisms. Therefore, they have been employed in various pharmaceutical applications. Plant peptidases have attracted considerable attention in various areas due to their specificity, stability across a diverse range of pH and temperatures, and safety profile. Here, we have focused on the use of plant peptidases, mostly papain and bromelain, to produce biologically active peptides, which confer various health advantages, including antioxidant, antimicrobial, antihypertensive, analgesic, antidiabetic, and anti-inflammatory effects. We have also discussed the importance of the action mechanism of peptidases for generating bioactive peptides with specific sequences and functions, the ecological and sustainability benefits of plant-derived peptidases compared to animal alternatives, digestive stability and bioavailability of peptides, as well as some obstacles to the commercialization of bioactive peptides and key challenges in peptidase-based industrial applications. Finally, we have examined enzyme immobilization as a viable method to enhance the production of bioactive peptides, offering numerous advantages in both research and industry contexts.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-04-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144014341","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}

引用次数: 0

Expression, Purification, and Preliminary Characterization of Putative Protein Tyrosine Phosphatase Oca1. 蛋白酪氨酸磷酸酶Oca1的表达、纯化和初步鉴定。

IF 1 4区生物学

Protein and Peptide Letters Pub Date : 2025-04-10 DOI: 10.2174/0109298665388367250404095840

Madhu Patel, Ashutosh Kumar, Kam Y J Zhang, Md Sohail Akhtar

{"title":"Expression, Purification, and Preliminary Characterization of Putative Protein Tyrosine Phosphatase Oca1.","authors":"Madhu Patel, Ashutosh Kumar, Kam Y J Zhang, Md Sohail Akhtar","doi":"10.2174/0109298665388367250404095840","DOIUrl":"https://doi.org/10.2174/0109298665388367250404095840","url":null,"abstract":"Introduction/objective: Protein phosphatases act as counterparts to protein kinases and are considered crucial for the homeostatic balance of cell signalling. In contrast to kinases, which can be categorized according to their substrate specificity, phosphatases are versatile and can detect substrates with much less distinction; hence, it is challenging to identify the physiological phosphatase-substrate pair. The Oca1 of Saccharomyces cerevisiae is a putative protein tyrosine phosphatase (PTP) and is required for cell cycle arrest in response to oxidative stress. The Oca1 mutants are sensitive to mTOR inhibitors, such as caffeine and rapamycin, and are involved in the regulation of TOR function. In an earlier research work, the enzyme exhibited no in vitro phosphatase activity and it was suggested that post-translational modifications or additional factors are necessary for it to be functional.Methods: The modeling of Oca1 was performed to gain insight into the structural aspects. The full- length enzyme, as well as the enzyme without the N-terminal extension, was cloned, expressed, and purified to homogeneity. The structure, function, and stability of the purified enzyme were assessed using circular dichroism, fluorescence, and visible spectroscopy studies.Results: The Oca1 was expressed and purified from Escherichia coli. The enzyme has been found to be functional, stable, and exist in an extended monomeric form, with a molecular mass of about 27 kDa. The enzyme without the extended N-terminal random coil has also been functional and slightly more stable than the full-length Oca1.Conclusion: The purified functional enzyme may be used to gain insights into the biochemical aspects and its role in bioengineering.","PeriodicalId":20736,"journal":{"name":"Protein and Peptide Letters","volume":" ","pages":""},"PeriodicalIF":1.0,"publicationDate":"2025-04-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144029012","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}

引用次数: 0