Biochimie最新文献_第4页

Enhanced activity of split trehalase biosensors by interspecies domain combineering 通过种间结构域组合提高分离式三卤甲烷酶生物传感器的活性。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.09.013

Yongpeng Fu, Jeroen De Buck

{"title":"Enhanced activity of split trehalase biosensors by interspecies domain combineering","authors":"Yongpeng Fu, Jeroen De Buck","doi":"10.1016/j.biochi.2024.09.013","DOIUrl":"10.1016/j.biochi.2024.09.013","url":null,"abstract":"<div><div>The split trehalase biosensor has potential as a versatile diagnostic technology. Split enzymes are engineered proteins, divided into inactive fragments, which can reassemble and regain activity when brought together by an analyte. The split TreA biosensor requires no sample processing and produces stable signals (in the form of glucose). Split trehalase reagents can function in blood, but periplasmic trehalase of <em>E. coli</em> requires blood acidification for maximal activity. The objective of this study was to obtain split trehalase with near physiological pH optimum. For this purpose, periplasmic trehalases of <em>Cellvibrio</em> spp. with higher activity at neutral pH, were split in analogy with the <em>E</em>. <em>coli</em> TreA into hood and catalytic domains. However, these split trehalases displayed self-complementation due to spontaneous reassembly. In contrast, when catalytic domains of <em>Cellvibrio</em> trehalases were combined with <em>E. coli</em> hood domains, these hybrids displayed conditional complementation capacity when split trehalase fragments fused to immunoglobulin-binding protein G (STIGA) were used to quantify immunoglobulin concentrations. Other hybrid combinations of <em>Cellvibrio</em> spp. had increased activity compared to the cognate pairs, albeit with strong self-complementation. A mutagenesis analysis of residues responsible for self-complementation led to uncoupling of self-complementation from allostery. The Michaelis-Menten kinetics of <em>Cellvibrio</em> enzymes and fragment pairs confirmed improved activity of a mutated hybrid pair of <em>Cellvibrio</em> hood and catalytic domains at physiological pH. In conclusion, the improvements in pH optimum and activity, demonstrated with STIGA, can now be leveraged to enhance other variations of the split trehalase biosensor platform, broadening its utility for testing clinical samples.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"228 ","pages":"Pages 167-175"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142585151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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Dietary monoterpenoids and human health: Unlocking the potential for therapeutic use 膳食单萜与人类健康：挖掘治疗潜力。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.09.002

Barbora Vyhlídalová , Karolína Ondrová , Iveta Zůvalová

引用次数: 0

In silico-driven identification and experimental confirmation of antifungal proteins (AFPs) against Candida albicans 针对白色念珠菌的抗真菌蛋白（AFPs）的硅学鉴定和实验确认。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.08.007

Jyoti Sankar Prusty, Awanish Kumar

{"title":"In silico-driven identification and experimental confirmation of antifungal proteins (AFPs) against Candida albicans","authors":"Jyoti Sankar Prusty, Awanish Kumar","doi":"10.1016/j.biochi.2024.08.007","DOIUrl":"10.1016/j.biochi.2024.08.007","url":null,"abstract":"<div><div>Mycoses infect millions of people annually across the world. The most common mycosis agent, <em>Candida albicans</em> is responsible for a great deal of illness and death. <em>C. albicans</em> infection is becoming more widespread and the current antifungals polyenes, triazoles, and echinocandins are less efficient against it. Investigating antifungal peptides (AFPs) as therapeutic is gaining momentum. Therefore, we used MALDI-TOF/MS analysis to identify AFPs and protein-protein docking to analyze their interactions with the <em>C. albicans</em> target protein. Some microorganisms with strong antifungal action against <em>C</em>. <em>albicans</em> were selected for the isolation of AFPs. Using MALDI-TOF/MS, we identified 3 AFPs Chitin binding protein (ACW83017.1; <em>Bacillus licheniformis</em>), the bifunctional protein GlmU (BBQ13478.1; <em>Stenotrophomonas maltophilia</em>), and zinc metalloproteinase aureolysin (BBA25172.1; Staphylococcus aureus). These AFPs showed robust interactions with <em>C. albicans</em> target protein Sap5. We deciphered some important residues in identified APFs and highlighted interaction with Sap5 through hydrogen bonds, protein-protein interactions, and salt bridges using protein-protein docking and MD simulations. The three discovered AFPs-Sap5 complexes exhibit different levels of stability, as seen by the RMSD analysis and interaction patterns. Among protein-protein interactions, the remarkable stability of the BBQ25172.1-2QZX complex highlights the role of salt bridges and hydrogen bonds. Identified AFPs could be further studied for developing successful antifungal candidates and peptide-based new antifungal therapeutic strategies as fresh insights into addressing antifungal resistance also.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"228 ","pages":"Pages 44-57"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141972460","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

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Inside front cover-EDB

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/S0300-9084(24)00303-1

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Insight into the transcriptional regulation of key genes involved in proline metabolism in plants under osmotic stress 渗透胁迫下植物脯氨酸代谢关键基因转录调控的深入研究

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.08.006

Shengjie Yan, Meng Zhan, Zhi Liu, Xianwen Zhang

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Novel chimeric peptides based on endomorphins and ghrelin receptor antagonist produced supraspinal antinociceptive effects with reduced acute tolerance in mice 基于内啡肽和胃泌素受体拮抗剂的新型嵌合肽可产生脊髓上部抗痛觉效应，并降低小鼠的急性耐受性。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.08.010

Bing Wu , Songxia Cheng , Fuyan Liu , Jia Wei , Yongling Liu , Teng Qian , Jiali Ding , Biao Xu , Jie Wei

{"title":"Novel chimeric peptides based on endomorphins and ghrelin receptor antagonist produced supraspinal antinociceptive effects with reduced acute tolerance in mice","authors":"Bing Wu , Songxia Cheng , Fuyan Liu , Jia Wei , Yongling Liu , Teng Qian , Jiali Ding , Biao Xu , Jie Wei","doi":"10.1016/j.biochi.2024.08.010","DOIUrl":"10.1016/j.biochi.2024.08.010","url":null,"abstract":"<div><div>It is widely recognized that developing bi- or multifunctional opioid compounds could offer a valuable approach to pain management with fewer side effects compared to single-target compounds. In this study, we designed and characterized two novel chimeric peptides, EM-1-DLS and EM-2-DLS, incorporating endomorphins (EMs) and the ghrelin receptor antagonist [D-Lys3]-GHRP-6 (DLS). Functional assays demonstrated that EM-1-DLS and EM-2-DLS acted as κ-opioid receptor (κ-OR)-preferring agonists, weak μ-opioid receptors (μ-OR) and ghrelin receptor (GHSR) agonists. Upon intracerebroventricular (i.c.v.) administration in mice, both EM-1-DLS and EM-2-DLS exhibited dose- and time-dependent antinociceptive effects in the tail withdrawal test. EM-1-DLS demonstrated the highest antinociceptive potency among the peptides, with an ED<sub>50</sub> approximately 8-fold greater than EM-1, while EM-2-DLS showed comparable effects to EM-2. The antinociceptive actions of EM-1-DLS involved activation of GHS-R1α, μ-OR, and κ-OR, whereas EM-2-DLS acted via GHS-R1α, δ-OR, and κ-OR pathways. Additionally, acute antinociceptive tolerance was investigated, revealing that EM-1-DLS induced a tolerance ratio of 2.33-fold, significantly lower than the 5.19-fold ratio induced by EM-1. Cross-tolerance ratios between the chimeric peptides and EMs ranged from 0.92 to 1.76, indicating reduced tolerance compared to EMs alone. These findings highlight the potential of these chimeric peptides to mitigate pain with diminished tolerance development, suggesting a promising strategy for the development of new analgesic therapies with improved safety profiles.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"228 ","pages":"Pages 58-70"},"PeriodicalIF":3.3,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"141989769","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

Orchestra of ligand-activated transcription factors in the molecular symphony of SERPINE 1 / PAI-1 gene regulation 配体激活的转录因子在 SERPINE 1 / PAI-1 基因调控的分子交响乐中的乐队。

IF 3.3 3区生物学

Biochimie Pub Date : 2025-01-01 DOI: 10.1016/j.biochi.2024.09.010

Aneta Vrzalova, Radim Vrzal

引用次数: 0

Revealing O-acetylhomoserine sulfhydrylase involved in direct sulfhydrylation pathway in Clostridium tetani 揭示破伤风梭菌参与直接巯基化途径的o -乙酰纯丝氨酸巯基化酶。

IF 3.3 3区生物学

Biochimie Pub Date : 2024-12-28 DOI: 10.1016/j.biochi.2024.12.014

Vitalia V. Kulikova , Natalya V. Anufrieva , Elena A. Morozova , Marat M. Khisamov , Yaroslav V. Tkachev , Mikhail I. Kotlov , Yury F. Belyi , Vasiliy S. Koval , Svetlana V. Revtovich , Pavel N. Solyev

{"title":"Revealing O-acetylhomoserine sulfhydrylase involved in direct sulfhydrylation pathway in Clostridium tetani","authors":"Vitalia V. Kulikova , Natalya V. Anufrieva , Elena A. Morozova , Marat M. Khisamov , Yaroslav V. Tkachev , Mikhail I. Kotlov , Yury F. Belyi , Vasiliy S. Koval , Svetlana V. Revtovich , Pavel N. Solyev","doi":"10.1016/j.biochi.2024.12.014","DOIUrl":"10.1016/j.biochi.2024.12.014","url":null,"abstract":"<div><div>Bacterial methionine biosynthesis is an attractive target for research due to its central role in cellular metabolism, as most steps of this pathway are missing in mammals. Up to now little is known about sulfur metabolism in pathogenic <em>Clostridia</em> species, making the study of the enzymes of Cys/Met metabolism in <em>Clostridium tetani</em> particularly relevant. Analysis of the <em>C. tetani</em> genome has shown that the bacterium is capable of synthesizing methionine by direct sulfhydration. In this study, we describe purification of recombinant <em>O</em>-acetylhomoserine sulfhydrylase, a member of the Cys/Met metabolism pyridoxal 5′-phosphate-dependent enzyme family, from <em>C. tetani</em> for the first time. The gene encoding <em>O</em>-acetylhomoserine sulfhydrylase was cloned into the pET-28a(+) vector and expressed in <em>Escherichia coli</em>. The expression product was purified and identified as a 462-amino acid protein with a molecular mass of ∼50 kDa as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The deduced amino acid sequence of the <em>C. tetani</em> enzyme showed a high degree of similarity to <em>O</em>-acetylhomoserine sulfhydrylases from other bacterial sources. We confirmed the <em>O</em>-acetylhomoserine sulfhydrylase activity, and found the enzyme to be optimally active at pH 7.5 and 50 °C. The native enzyme assembles into a homotetramer of approx. 200 kDa as revealed by gel filtration. The obtained enzyme is capable of <span>l</span>-methionine formation using methanethiol as a sulfur source, that has been revealed by <sup>1</sup>H NMR spectral data. These findings broaden the understanding of the role of <em>O</em>-acetylhomoserine sulfhydrylase in <em>C. tetani</em> Cys/Met metabolism and provide a basis for its future investigations and research.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"231 ","pages":"Pages 146-154"},"PeriodicalIF":3.3,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142908042","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"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

The cysteine protease legumain decreases glucose metabolism and enhances fatty acid uptake in human myotubes 半胱氨酸蛋白酶豆类降低葡萄糖代谢和提高脂肪酸摄取在人肌管。

IF 3.3 3区生物学

Biochimie Pub Date : 2024-12-28 DOI: 10.1016/j.biochi.2024.12.012

Nimo Mukhtar Mohamud Osoble , Ngoc Nguyen Lunde , Abbas Jafari , G. Hege Thoresen , Rigmor Solberg , Arild C. Rustan

{"title":"The cysteine protease legumain decreases glucose metabolism and enhances fatty acid uptake in human myotubes","authors":"Nimo Mukhtar Mohamud Osoble , Ngoc Nguyen Lunde , Abbas Jafari , G. Hege Thoresen , Rigmor Solberg , Arild C. Rustan","doi":"10.1016/j.biochi.2024.12.012","DOIUrl":"10.1016/j.biochi.2024.12.012","url":null,"abstract":"<div><div>Skeletal muscle has an important role in whole body energy metabolism and various proteases are involved in skeletal muscle functions. We have previously identified the cysteine protease legumain in cultured human skeletal muscle cells. However, the potential role of legumain in regulation of energy metabolism remains unexplored. This study aimed to investigate cellular uptake, processing, and activation of prolegumain in human myotubes. Additionally, we sought to determine the effects of prolegumain on energy substrate metabolism in these cells. During differentiation of human myoblast to myotubes, legumain mRNA expression and activity were upregulated. Interestingly, legumain activity in myotubes was inversely correlated with the body mass index (BMI) of the obese cell donors. Myotubes exposed to conditioned medium enriched in prolegumain during the last two days of differentiation demonstrated the capacity to internalize and process prolegumain into its active form. Pre-treatment with prolegumain induced a metabolic shift towards increased fatty acid uptake in myotubes, as evidenced by elevated oleic acid uptake whereas glucose uptake and oxidation were reduced. The metabolic changes were not reversed by a legumain inhibitor, indicating a different mechanism for this effect. The metabolic alterations were accompanied by increased mRNA expression of the fatty acid transporter CD36, whereas the glucose transporter GLUT1 mRNA level remained unchanged. These findings suggest that legumain may play a regulatory role in skeletal muscle energy metabolism, highlighting its potential as a novel therapeutic target of metabolic disorders.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"231 ","pages":"Pages 127-136"},"PeriodicalIF":3.3,"publicationDate":"2024-12-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142908045","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

The neutralisation and inhibitory effects of new tRNA-mimetic aptamer against cytotoxin-induced cytotoxicity in human skin keratinocytes

IF 3.3 3区生物学

Biochimie Pub Date : 2024-12-25 DOI: 10.1016/j.biochi.2024.12.011

Jia Jin Hiu, Hock Siew Tan, Michelle Khai Khun Yap

{"title":"The neutralisation and inhibitory effects of new tRNA-mimetic aptamer against cytotoxin-induced cytotoxicity in human skin keratinocytes","authors":"Jia Jin Hiu, Hock Siew Tan, Michelle Khai Khun Yap","doi":"10.1016/j.biochi.2024.12.011","DOIUrl":"10.1016/j.biochi.2024.12.011","url":null,"abstract":"<div><div>Cytotoxin (CTX) is the major venom toxin in cobra venom, accounting for the dermonecrosis in cobra envenomed patients. Conventional antibody-based antivenoms have limited efficacy in the treatment for CTX-manifested dermonecrosis, highlighting the urgent needs for discovery of new therapeutics. This study aims to discover oligonucleotide-based aptamers which target the functional epitopes of CTX to inhibit its cytotoxicity. Genomic tRNA Database (GtRNAdb) was acquired for the computational selection and design of potential tRNA-mimetic aptamers (AptRNAs) targeting CTX. The binding characteristics of the AptRNAs were further validated by electromobility shift assay (EMSA), and enzyme-linked oligonucleotide assay (ELONA). The best candidate was also examined for neutralisation potency and efficacy. The computational analyses identified six consensus AptRNA clusters, whereby three AptRNAs showed significant binding for CTX with an ascending ranking from AptRNA4 > AptRNA6 > AptRNA5. These AptRNAs bind to the functional epitopes of CTX. AptRNA6 had the highest binding specificity towards CTX, with a binding affinity (K<sub>D</sub>) of 14.45 nM. Furthermore, AptRNA6 exhibited the best neutralisation and rescue potency in CTX-induced cytotoxicity. Even at low treatment concentrations, AptRNA6 also demonstrated the highest potency in rescuing experimental post-envenomed model by cobra venoms of medically important species. Overall, the findings concluded the potential of AptRNA6 as synthetic biotherapeutic against cobra venom CTX.</div></div>","PeriodicalId":251,"journal":{"name":"Biochimie","volume":"231 ","pages":"Pages 116-126"},"PeriodicalIF":3.3,"publicationDate":"2024-12-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143592469","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0