Pub Date : 2023-10-01DOI: 10.1016/j.cbpa.2023.102375
Delilah Jewel , Quan Pham , Abhishek Chatterjee
Directed evolution is a powerful technique that uses principles of natural evolution to enable the development of biomolecules with novel functions. However, the slow pace of natural evolution does not support the demand for rapidly generating new biomolecular functions in the laboratory. Viruses offer a unique path to design fast laboratory evolution experiments, owing to their innate ability to evolve much more rapidly than most living organisms, facilitated by a smaller genome size that tolerate a high frequency of mutations, as well as a fast rate of replication. These attributes offer a great opportunity to evolve various biomolecules by linking their activity to the replication of a suitable virus. This review highlights the recent advances in the application of virus-assisted directed evolution of designer biomolecules in both prokaryotic and eukaryotic cells.
{"title":"Virus-assisted directed evolution of biomolecules","authors":"Delilah Jewel , Quan Pham , Abhishek Chatterjee","doi":"10.1016/j.cbpa.2023.102375","DOIUrl":"10.1016/j.cbpa.2023.102375","url":null,"abstract":"<div><p>Directed evolution is a powerful technique that uses principles of natural evolution to enable the development of biomolecules with novel functions. However, the slow pace of natural evolution does not support the demand for rapidly generating new biomolecular functions in the laboratory. Viruses offer a unique path to design fast laboratory evolution experiments, owing to their innate ability to evolve much more rapidly than most living organisms, facilitated by a smaller genome size that tolerate a high frequency of mutations, as well as a fast rate of replication. These attributes offer a great opportunity to evolve various biomolecules by linking their activity to the replication of a suitable virus. This review highlights the recent advances in the application of virus-assisted directed evolution of designer biomolecules in both prokaryotic and eukaryotic cells.</p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9994988","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Protein-protein and protein-RNA interactions are essential for cell function and survival. These interactions facilitate the formation of ribonucleoprotein complexes and biomolecular condensates via phase separation. Such assembly is involved in transcription, splicing, translation and stress response. When dysregulated, proteins and RNA can undergo irreversible aggregation which can be cytotoxic and pathogenic. Despite technical advances in investigating biomolecular condensates, achieving the necessary spatiotemporal resolution to deduce the parameters that govern their assembly and behavior has been challenging. Many laboratories have applied advanced microscopy methods for imaging condensates. For example, single molecule imaging methods have enabled the detection of RNA-protein interaction, protein-protein interaction, protein conformational dynamics, and diffusional motion of molecules that report on the intrinsic molecular interactions underlying liquid-liquid phase separation. This review will outline advances in both microscopy and spectroscopy techniques which allow single molecule detection and imaging, and how these techniques can be used to probe unique aspects of biomolecular condensates.
{"title":"Advanced imaging techniques for studying protein phase separation in living cells and at single-molecule level","authors":"Gemechu Mekonnen , Nathalie Djaja , Xincheng Yuan , Sua Myong","doi":"10.1016/j.cbpa.2023.102371","DOIUrl":"10.1016/j.cbpa.2023.102371","url":null,"abstract":"<div><p><span>Protein-protein and protein-RNA interactions are essential for cell function and survival. These interactions facilitate the formation of ribonucleoprotein<span> complexes and biomolecular condensates via phase separation. Such assembly is involved in transcription, splicing, translation and stress response. When dysregulated, proteins and RNA can undergo irreversible aggregation which can be cytotoxic and pathogenic. Despite technical advances in investigating biomolecular condensates, achieving the necessary spatiotemporal resolution to deduce the parameters that govern their assembly and behavior has been challenging. Many laboratories have applied advanced </span></span>microscopy<span><span> methods for imaging condensates. For example, single molecule imaging<span> methods have enabled the detection of RNA-protein interaction, protein-protein interaction, protein conformational dynamics, and diffusional motion of molecules that report on the intrinsic molecular interactions<span> underlying liquid-liquid phase separation. This review will outline advances in both microscopy and spectroscopy techniques which allow </span></span></span>single molecule detection and imaging, and how these techniques can be used to probe unique aspects of biomolecular condensates.</span></p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10038502","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-01DOI: 10.1016/j.cbpa.2023.102377
Xiaoyan Chen , Bo Li
Living organisms have evolved various strategies to incorporate sulfur and selenium into bioactive natural products. These chalcogen-containing compounds serve important and diverse biological functions for their producers and many of them are essential medicines against infectious diseases and cancer. We review recent advances in the biosynthesis of some sulfur/selenium-containing natural products with a focus on the formation or cleavage of C–S/C–Se bonds. We highlight unusual enzymes that catalyze these transformations, describe their proposed mechanisms, and discuss how understanding these enzymes may facilitate the discovery and synthesis of novel natural products containing sulfur or selenium.
{"title":"How nature incorporates sulfur and selenium into bioactive natural products","authors":"Xiaoyan Chen , Bo Li","doi":"10.1016/j.cbpa.2023.102377","DOIUrl":"10.1016/j.cbpa.2023.102377","url":null,"abstract":"<div><p><span>Living organisms have evolved various strategies to incorporate sulfur and selenium into bioactive natural products. These chalcogen-containing compounds serve important and diverse biological functions for their producers and many of them are </span>essential medicines<span><span> against infectious diseases and cancer. We review recent advances in the biosynthesis of some sulfur/selenium-containing natural products with a focus on the formation or cleavage of C–S/C–Se bonds. We highlight unusual </span>enzymes that catalyze these transformations, describe their proposed mechanisms, and discuss how understanding these enzymes may facilitate the discovery and synthesis of novel natural products containing sulfur or selenium.</span></p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10031056","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-10-01DOI: 10.1016/j.cbpa.2023.102376
Teresa E. Sullivan , Servando Hernandez Vargas , Sukhen C. Ghosh , Solmaz AghaAmiri , Naruhiko Ikoma , Ali Azhdarinia
Cancer imaging is a rapidly evolving field due to the discovery of novel molecular targets and the availability of corresponding techniques to detect them with high precision, accuracy, and sensitivity. Nuclear medicine is the most widely used molecular imaging modality and has a growing toolkit of clinically used radiopharmaceuticals that enable whole-body tumor visualization, staging, and treatment monitoring for a variety of tumors in a non-invasive manner. The need for similar imaging capabilities in the operating room has led to the emergence of fluorescence-guided surgery (FGS) as a powerful technique that gives surgeons unprecedented ability to distinguish tumors from healthy tissues. While a variety of strategies have been used to develop contrast agents for FGS, the use of radiopharmaceuticals as models brings exceptional translational potential and has increasingly been explored. Here, we review strategies used to convert clinically used radiopharmaceuticals into fluorescent and multimodal counterparts. Unique preclinical and clinical capabilities stemming from radiopharmaceutical-based agent design are also discussed to illustrate the advantages of this approach.
{"title":"A translational blueprint for developing intraoperative imaging agents via radiopharmaceutical-guided drug design","authors":"Teresa E. Sullivan , Servando Hernandez Vargas , Sukhen C. Ghosh , Solmaz AghaAmiri , Naruhiko Ikoma , Ali Azhdarinia","doi":"10.1016/j.cbpa.2023.102376","DOIUrl":"10.1016/j.cbpa.2023.102376","url":null,"abstract":"<div><p>Cancer imaging is a rapidly evolving field due to the discovery of novel molecular targets and the availability of corresponding techniques to detect them with high precision, accuracy, and sensitivity. Nuclear medicine is the most widely used molecular imaging<span> modality and has a growing toolkit of clinically used radiopharmaceuticals that enable whole-body tumor visualization, staging, and treatment monitoring for a variety of tumors in a non-invasive manner. The need for similar imaging capabilities in the operating room has led to the emergence of fluorescence-guided surgery (FGS) as a powerful technique that gives surgeons unprecedented ability to distinguish tumors from healthy tissues. While a variety of strategies have been used to develop contrast agents for FGS, the use of radiopharmaceuticals as models brings exceptional translational potential and has increasingly been explored. Here, we review strategies used to convert clinically used radiopharmaceuticals into fluorescent and multimodal counterparts. Unique preclinical and clinical capabilities stemming from radiopharmaceutical-based agent design are also discussed to illustrate the advantages of this approach.</span></p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9979371","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-09-28DOI: 10.1016/j.cbpa.2023.102389
Evan R. Stair, Leslie M. Hicks
The post-translational modification of cysteine to diverse oxidative states is understood as a critical cellular mechanism to combat oxidative stress. To study the role of cysteine oxidation, cysteine enrichments and subsequent analysis via mass spectrometry are necessary. As such, technologies and methods are rapidly developing for sensitive and efficient enrichments of cysteines to further explore its role in signaling pathways. In this review, we analyze recent developments in methods to miniaturize cysteine enrichments, analyze the underexplored disulfide bound redoxome, and quantify site-specific cysteine oxidation. We predict that further development of these methods will improve cysteine coverage across more diverse organisms than those previously studied and elicit novel roles cysteines play in stress response.
{"title":"Recent advances in mass spectrometry-based methods to investigate reversible cysteine oxidation","authors":"Evan R. Stair, Leslie M. Hicks","doi":"10.1016/j.cbpa.2023.102389","DOIUrl":"10.1016/j.cbpa.2023.102389","url":null,"abstract":"<div><p><span><span>The post-translational modification of cysteine to diverse oxidative states is understood as a critical cellular mechanism to combat oxidative stress. To study the role of cysteine </span>oxidation, cysteine enrichments and subsequent analysis via mass spectrometry are necessary. As such, technologies and methods are rapidly developing for sensitive and efficient enrichments of cysteines to further explore its role in </span>signaling pathways<span>. In this review, we analyze recent developments in methods to miniaturize cysteine enrichments, analyze the underexplored disulfide bound redoxome, and quantify site-specific cysteine oxidation. We predict that further development of these methods will improve cysteine coverage across more diverse organisms than those previously studied and elicit novel roles cysteines play in stress response.</span></p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-09-28","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"41096617","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-09-14DOI: 10.1016/j.cbpa.2023.102387
Xiufang Wang , Guozhi Xiao
Here, chemical syntheses of long, branched and complex glycans over 10-mer from plants are summarized, which highlights amylopectin 20-mer from starch, 17-mer from carthamus tinctorius, α-glucan 30-mer from Longan, 19-mer from psidium guajava and 11-mer from dendrobium huoshanense. The glycans assembly strategies, protecting groups utilization and glycosylation methods discussed here will inspire the efficient synthesis of diverse complex glycans with many 1,2-cis glycosidic linkages.
{"title":"Recent chemical synthesis of plant polysaccharides","authors":"Xiufang Wang , Guozhi Xiao","doi":"10.1016/j.cbpa.2023.102387","DOIUrl":"10.1016/j.cbpa.2023.102387","url":null,"abstract":"<div><p><span>Here, chemical syntheses of long, branched and complex glycans<span> over 10-mer from plants are summarized, which highlights amylopectin 20-mer from starch, 17-mer from </span></span><span><em>carthamus tinctorius</em></span>, α-glucan 30-mer from <span><em>Longan</em></span>, 19-mer from <span><em>psidium</em><em> guajava</em></span> and 11-mer from <span><em>dendrobium</em><em> huoshanense</em></span><span>. The glycans assembly strategies, protecting groups utilization and glycosylation methods discussed here will inspire the efficient synthesis of diverse complex glycans with many 1,2-</span><em>cis</em><span> glycosidic linkages.</span></p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-09-14","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"10268661","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-09-04DOI: 10.1016/j.cbpa.2023.102386
Cong Liu, Xin Zhang
{"title":"Navigating the terrain of protein aggregation and phase separation - A chemical biology perspective","authors":"Cong Liu, Xin Zhang","doi":"10.1016/j.cbpa.2023.102386","DOIUrl":"https://doi.org/10.1016/j.cbpa.2023.102386","url":null,"abstract":"","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-09-04","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"1892563","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-08-01DOI: 10.1016/j.cbpa.2023.102316
Christopher R. Mansfield , Michael E. Chirgwin , Emily R. Derbyshire
Intracellular protozoan parasites are responsible for wide-spread infectious diseases. These unicellular pathogens have complex, multi-host life cycles, which present challenges for investigating their basic biology and for discovering vulnerabilities that could be exploited for disease control. Throughout development, parasite proteomes are dynamic and support stage-specific functions, but detection of these proteins is often technically challenging and complicated by the abundance of host proteins. Thus, to elucidate key parasite processes and host–pathogen interactions, labeling strategies are required to track pathogen proteins during infection. Herein, we discuss the application of bioorthogonal non-canonical amino acid tagging and proximity-dependent labeling to broadly study protozoan parasites and include outlooks for future applications to study Plasmodium, the causative agent of malaria. We highlight the potential of these technologies to provide spatiotemporal labeling with selective parasite protein enrichment, which could enable previously unattainable insight into the biology of elusive developmental stages.
{"title":"Labeling strategies to track protozoan parasite proteome dynamics","authors":"Christopher R. Mansfield , Michael E. Chirgwin , Emily R. Derbyshire","doi":"10.1016/j.cbpa.2023.102316","DOIUrl":"https://doi.org/10.1016/j.cbpa.2023.102316","url":null,"abstract":"<div><p><span>Intracellular protozoan parasites are responsible for wide-spread infectious diseases. These unicellular pathogens have complex, multi-host life cycles, which present challenges for investigating their basic biology and for discovering vulnerabilities that could be exploited for disease control. Throughout development, parasite proteomes are dynamic and support stage-specific functions, but detection of these proteins is often technically challenging and complicated by the abundance of host proteins. Thus, to elucidate key parasite processes and host–pathogen interactions, labeling strategies are required to track pathogen proteins during infection. Herein, we discuss the application of bioorthogonal non-canonical amino acid tagging and proximity-dependent labeling to broadly study protozoan parasites and include outlooks for future applications to study </span><em>Plasmodium,</em> the causative agent of malaria. We highlight the potential of these technologies to provide spatiotemporal labeling with selective parasite protein enrichment, which could enable previously unattainable insight into the biology of elusive developmental stages.</p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"1527688","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-08-01DOI: 10.1016/j.cbpa.2023.102322
Daniel G. Dumitrescu , Stavroula K. Hatzios
Low-molecular-weight (LMW) thiols are an abundant class of cysteine-derived small molecules found in all forms of life that maintain reducing conditions within cells. While their contributions to cellular redox homeostasis are well established, LMW thiols can also mediate other aspects of cellular physiology, including intercellular interactions between microbial and host cells. Here we discuss emerging roles for these redox-active metabolites at the host–microbe interface. We begin by providing an overview of chemical and computational approaches to LMW-thiol discovery. Next, we highlight mechanisms of virulence regulation by LMW thiols in infected cells. Finally, we describe how microbial metabolism of these compounds may influence host physiology.
{"title":"Emerging roles of low-molecular-weight thiols at the host–microbe interface","authors":"Daniel G. Dumitrescu , Stavroula K. Hatzios","doi":"10.1016/j.cbpa.2023.102322","DOIUrl":"https://doi.org/10.1016/j.cbpa.2023.102322","url":null,"abstract":"<div><p><span>Low-molecular-weight (LMW) thiols are an abundant class of cysteine-derived small molecules<span> found in all forms of life that maintain reducing conditions within cells. While their contributions to cellular redox homeostasis are well established, LMW thiols can also mediate other aspects of cellular physiology, including intercellular interactions between microbial and host cells. Here we discuss emerging roles for these redox-active metabolites at the host–microbe interface. We begin by providing an overview of chemical and computational approaches to LMW-thiol discovery. Next, we highlight mechanisms of virulence regulation by LMW thiols in infected cells. Finally, we describe how </span></span>microbial metabolism of these compounds may influence host physiology.</p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"3451952","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pub Date : 2023-08-01DOI: 10.1016/j.cbpa.2023.102306
Richard J. Spears, Vijay Chudasama
Advances in the site-specific chemical modification of proteins, also referred to as protein bioconjugation, have proved instrumental in revolutionary approaches to designing new protein-based therapeutics. Of the sites available for protein modification, cysteine residues or the termini of proteins have proved especially popular owing to their favorable properties for site-specific modification. Strategies that, therefore, specifically target cysteine at the termini offer a combination of these favorable properties of cysteine and termini bioconjugation. In this review, we discuss these strategies with a particular focus on those reported recently and provide our opinion on the future direction of the field.
{"title":"Recent advances in N- and C-terminus cysteine protein bioconjugation","authors":"Richard J. Spears, Vijay Chudasama","doi":"10.1016/j.cbpa.2023.102306","DOIUrl":"https://doi.org/10.1016/j.cbpa.2023.102306","url":null,"abstract":"<div><p>Advances in the site-specific chemical modification of proteins, also referred to as protein bioconjugation, have proved instrumental in revolutionary approaches to designing new protein-based therapeutics. Of the sites available for protein modification, cysteine residues or the termini of proteins have proved especially popular owing to their favorable properties for site-specific modification. Strategies that, therefore, specifically target cysteine at the termini offer a combination of these favorable properties of cysteine and termini bioconjugation. In this review, we discuss these strategies with a particular focus on those reported recently and provide our opinion on the future direction of the field.</p></div>","PeriodicalId":291,"journal":{"name":"Current Opinion in Chemical Biology","volume":null,"pages":null},"PeriodicalIF":7.8,"publicationDate":"2023-08-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"3451953","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}