Review
Biochemistry & Molecular Biology
Stephanie Sauve, Joseph Williamson, Adithya Polasa, Mahmoud Moradi
Summary: The major facilitator superfamily (MFS) of transporters consists of three classes of membrane transporters: symporters, uniporters, and antiporters. The conformational changes within the distinct transport cycles of MFS transporters are believed to follow the rocker-switch mechanism. This review compares the similarities and differences in conformational dynamics between symporters, uniporters, and antiporters of the MFS family using experimental and computational structural data.
Article
Microbiology
Ashley N. Williams, John Stavrinides
Summary: Pantoea Natural Product 3 (PNP-3) is an effective antibiotic against multi-drug resistant bacteria. The production and resistance of PNP-3 are regulated by two transporters, pnp3a and pnp3c. Disruption of pnp3a leads to impaired production and resistance, while disruption of pnp3c only reduces production. These findings provide important insights into the synthesis and resistance mechanism of PNP-3.
RESEARCH IN MICROBIOLOGY
(2022)
Review
Infectious Diseases
Jerusha Stephen, Fathima Salam, Manjusha Lekshmi, Sanath H. Kumar, Manuel F. Varela
Summary: This review article discusses the S. aureus-specific MFS multidrug efflux pump systems from a molecular mechanistic perspective, including structure-function relationships, modulation of antimicrobial resistance mediated by MFS drug efflux pumps, and direction for future investigation.
Review
Biochemistry & Molecular Biology
Manuel F. Varela, Jerusha Stephen, Deeksha Bharti, Manjusha Lekshmi, Sanath Kumar
Summary: Bacterial pathogens resistant to multiple antimicrobial agents through active efflux pose a serious threat to public health. This review discusses recent molecular studies focused on modulating the antimicrobial efflux transporters to restore the clinical efficacy of infectious disease chemotherapy.
Article
Medicine, Research & Experimental
Angelika Janaszkiewicz, Agota Toth, Quentin Faucher, Helene Arnion, Nicolas Vedrenne, Chantal Barin-Le Guellec, Pierre Marquet, Florent Di Meo
Summary: The Organic Anion Transporter 1 (hOAT1) is a membrane transporter involved in drug elimination by the kidney. This study aimed to investigate fragments of hOAT1 structure and function in order to better understand its transport cycle. Molecular dynamics simulations suggested two potential binding sites for the substrate adefovir, including a new inner binding cavity. The binding modes of the co-substrate alpha-ketoglutarate were also explored. The findings provide insights into the transport impairments caused by hOAT1 mutations.
BIOMEDICINE & PHARMACOTHERAPY
(2023)
Article
Multidisciplinary Sciences
Yu Liu, Chenghan Li, Meghna Gupta, Nidhi Verma, Atul Kumar Johri, Robert M. Stroud, Gregory A. Voth
Summary: Phosphate is a crucial metabolite in cells, and proton-coupled phosphate transporters play important roles in phosphate uptake in plants, fungi, and certain pathogenic protozoa. Research has shown that deprotonation of D324 triggers phosphate release in a eukaryotic phosphate transporter, providing insights into the proton-driven phosphate transport mechanism.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2021)
Article
Microbiology
Jennifer C. Chang, Reid Wilkening, Kate M. Rahbari, Michael J. Federle
Summary: Cell-cell signaling mediated by Rgg-family transcription factors and their cognate pheromones plays a crucial role in regulating various phenotypes in Firmicutes. In particular, the Rgg2/3 quorum sensing system in Streptococcus pyogenes has been shown to influence the expression of secreted virulence factors and antibiotic susceptibility, with dysregulation by the deletion of an Rgg2/3 QS-regulated MFS transporter. This study highlights the complex regulatory mechanisms involved in the expression of virulence factors and antibiotic sensitivity in bacterial pathogens.
JOURNAL OF BACTERIOLOGY
(2022)
Article
Microbiology
Wenzhe Tian, Jiayang Qin, Congcong Lian, Qingshou Yao, Xiuwen Wang
Summary: This study identified a mutant gene mfs coding for the major facilitator superfamily (MFS) protein in Bacillus coagulans Na-2, which has better resistance to sodium lactate stress. Expression of MFS-2-6 and MFS-Na-2 improved L-LA production at low pH, with MFS-Na-2 showing better performance likely due to more appropriate intracellular pH during fermentation. The MFS protein identified here has potential application in environmentally friendly L-LA production.
Article
Genetics & Heredity
Jian Diao, Shuxuan Li, Ling Ma, Ping Zhang, Jianyang Bai, Jiaqi Wang, Xiaoqian Ma, Wei Ma
Summary: This study systematically analyzed the structure and function of genes and proteins in the PtrMFS family, indicating that poplar MFS genes play key roles in plant development and response to biological stressors.
FRONTIERS IN GENETICS
(2021)
Article
Chemistry, Multidisciplinary
Ashwini Nawade, Kumar Babu Busi, Kunchanapalli Ramya, Goutam Kumar Dalapati, Sabyasachi Mukhopadhyay, Sabyasachi Chakrabortty
Summary: Proteins are essential and highly complex substances in living organisms with biocompatible properties, and their electrical conductivity can be modulated by incorporating metal nanodusters. This research has potential applications for preparing biocompatible electronic devices.
Article
Biochemistry & Molecular Biology
Na Liu, Qiannan Wang, Chaozu He, Bang An
Summary: The study characterizes an MFS protein CgMFS1 in Colletotrichum gloeosporioides, showing its involvement in sugar transport, response to oxidative stress, and pathogenicity of the fungus.
CURRENT ISSUES IN MOLECULAR BIOLOGY
(2021)
Article
Chemistry, Organic
Huan Yue, Liangcheng Du
Summary: This study reveals the important role of hsaf-orf1 in the regulation and biosynthesis of HSAF and its analogs by controlling the import of sugars, which induce antifungal production.
Article
Biochemistry & Molecular Biology
Lixin He, Jinxin Chen, Pinwei Deng, Shumei Huang, Pian Liu, Chanjuan Wang, Xinjian Huang, Yue Li, Boyu Chen, Dongni Shi, Yunyun Xiao, Xiangfu Chen, Ying Ouyang, Libing Song, Chuyong Lin
Summary: Cyst(e)ine plays a crucial role in the synthesis of glutathione and its storage in lysosomes helps cancer cells maintain redox homeostasis. Breast cancer cells upregulate MFSD12 to increase lysosomal cyst(e)ine storage, which is released to maintain GSH levels and buffer oxidative stress. mTORC1 regulates MFSD12 by phosphorylating residue T254, and this switch modulates lysosomal cyst(e)ine levels in response to oxidative stress, enhancing cell fitness. MFSD12 mutation inhibits its function and suppresses tumor progression, while its overexpression correlates with poor chemotherapy response and prognosis in breast cancer patients.
Article
Biochemistry & Molecular Biology
Artem Stetsenko, Pavlo Stehantsev, Natalia O. Dranenko, Mikhail S. Gelfand, Albert Guskov
Summary: The study reports the functional and structural analysis of a membrane protein CmaX from a pathogenic Pseudomonas aeruginosa bacterium, which has a signature motif deviating from the canonical one. Despite the difference in composition, CmaX shows no changes in substrate selectivity or transport, suggesting that deviations from the canonical motif can easily alter during evolution.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2021)
Review
Biology
Manuel F. Varela, Anely Ortiz-Alegria, Manjusha Lekshmi, Jerusha Stephen, Sanath Kumar
Summary: A phospholipid membrane acts as a barrier for the passage of molecules across living cells, but this barrier can be circumvented by transporter proteins. The major facilitator superfamily includes transporters found in all living organisms and plays important roles in various biological processes. The antiporter motif, known as motif C, is involved in the molecular mechanism of antimicrobial efflux.
Article
Multidisciplinary Sciences
Jeremy R. Keown, Zihan Zhu, Loic Carrique, Haitian Fan, Alexander P. Walker, Itziar Serna Martin, Els Pardon, Jan Steyaert, Ervin Fodor, Jonathan M. Grimes
Summary: Influenza A viruses cause significant burdens to healthcare systems through seasonal epidemics and global pandemics. The viral RNA-dependent RNA polymerase plays a central role in the replication cycle of influenza viruses and is a potential target for antiviral development. By characterizing the inhibitory effect of nanobodies on the 1918 pandemic influenza virus polymerase complex, sensitive sites interfering with polymerase activity in vitro were identified, suggesting them as effective targets for potential influenza antiviral development.
NATURE COMMUNICATIONS
(2022)
Meeting Abstract
Biochemistry & Molecular Biology
David Salom, Arum Wu, Christopher L. Sander, Els Pardon, Jan Steyaert, Philip D. Kiser, Krzysztof Palczewski
Article
Multidisciplinary Sciences
Kapil Goutam, Francesco S. Ielasi, Els Pardon, Jan Steyaert, Nicolas Reyes
Summary: This study provides molecular insights into the key conformations of NTCP transport cycle and the recognition mechanisms of HBV/HDV receptors. These findings are expected to contribute to the development of liver disease therapies targeting NTCP.
Article
Multidisciplinary Sciences
Finn P. Maloney, Jeremi Kuklewicz, Robin A. Corey, Yunchen Bi, Ruoya Ho, Lukasz Mateusiak, Els Pardon, Jan Steyaert, Phillip J. Stansfeld, Jochen Zimmer
Summary: In this study, we investigated the substrate binding and polymer synthesis of hyaluronan synthase (HAS) using cryo-electron microscopy structures, biochemical analyses, and molecular dynamics simulations. Our findings provide detailed insights into the transmembrane process of HAS, including substrate selection, polymerization, and regulation of hyaluronan synthesis. This research is important for understanding the biosynthesis of acidic extracellular polysaccharides and one of the most abundant glycosaminoglycans in the human body.
Article
Biochemistry & Molecular Biology
John J. Kelly, Dale Tranter, Els Pardon, Gamma Chi, Holger Kramer, Lotta Happonen, Kelly M. Knee, Jay M. Janz, Jan Steyaert, Christine Bulawa, Ville O. Paavilainen, Juha T. Huiskonen, Wyatt W. Yue
Summary: The integrity of a cell's proteome relies on the correct folding of polypeptides, which is facilitated by the chaperonin TCP-1 ring complex (TRiC). Structural studies have provided insights into the architecture and substrate recognition of TRiC, but the fate of substrates inside the TRiC chamber has remained unclear. In this study, cryo-EM was used to determine the structure of endogenous human TRiC with substrates and cochaperone at different folding stages. The findings revealed the positioning of already-folded regions of client proteins at the chamber wall, allowing unstructured regions to achieve their native fold. The substrates engaged with different sections of the chamber during the folding process, while the cochaperone PhLP2A acted as a molecular strut between the substrate and TRiC chamber. These structural snapshots contribute to our understanding of client protein folding within TRiC.
NATURE STRUCTURAL & MOLECULAR BIOLOGY
(2022)
Article
Multidisciplinary Sciences
Ranjan K. Singh, Ahmed Soliman, Giambattista Guaitoli, Eliza Stoermer, Felix von Zweydorf, Thomas Dal Maso, Asmaa Oun, Laura Van Rillaer, Sven H. Schmidt, Deep Chatterjee, Joshua A. David, Els Pardon, Thomas U. Schwartz, Stefan Knapp, Eileen J. Kennedy, Jan Steyaert, Friedrich W. Herberg, Arjan Kortholt, Christian Johannes Gloeckner, Wim Versees
Summary: Mutations in the LRRK2 gene are a leading cause of Parkinson's disease, while overactivation of LRRK2 is associated with idiopathic form of the disease. Researchers have identified and characterized nanobodies that can bind to different domains of LRRK2 and inhibit or activate its activity. These nanobodies act through an allosteric inhibitor mechanism and provide potential therapeutic strategies for Parkinson's disease.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Biochemistry & Molecular Biology
Maxime Killer, Giada Finocchio, Haydyn D. T. Mertens, Dmitri I. Svergun, Els Pardon, Jan Steyaert, Christian Loew
Summary: Proton-coupled Oligopeptide Transporters (POTs) mediate the uptake of short di- and tripeptides. This study determined the structure of DtpC and provided insights into its ligand specificity using cryogenic electron microscopy (cryo-EM). The research highlights the value of nanobodies for structure determination of low molecular weight integral membrane proteins.
FRONTIERS IN MOLECULAR BIOSCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Lorenzo Maso, Filippo Vascon, Monica Chinellato, Frederic Goormaghtigh, Pierangelo Bellio, Enrica Campagnaro, Laurence Van Melderen, Maria Ruzzene, Els Pardon, Alessandro Angelini, Giuseppe Celenza, Jan Steyaert, Donatella Tondi, Laura Cendron
Summary: Antimicrobial resistance poses a threat to eradicating infectious diseases and reducing the effectiveness of available treatments. The bacterial SOS pathway is a key mechanism leading to resistance, and inhibiting this pathway may delay the evolution of antimicrobial resistance. In this study, nanobodies that bind to and block the SOS response in Escherichia coli were identified, paving the way for the development of new-generation antibiotic adjuvants for treating bacterial infections.
Article
Chemistry, Multidisciplinary
Kevin Van Holsbeeck, Baptiste Fischer, Simon Gonzalez, Charlene Gadais, Wim Versees, Jose C. Martins, Charlotte Martin, Alexandre Wohlkoenig, Jan Steyaert, Steven Ballet
Summary: RAS proteins play a crucial role in regulating intracellular signaling networks and mutations that stabilize their active state are associated with cancer development. The study investigated the potential of developing peptide mimetics to modulate RAS signaling by mimicking the complementarity-determining region 3 (CDR3) of the regulatory guanine nucleotide exchange factor (GEF) son of sevenless 1 (SOS1). Through optimization and conformational rigidification, CDR3 mimetics with half of the maximal activation potential of Nanobody14 (Nb14) were obtained, demonstrating the feasibility of modulating protein-protein interactions through structural mimicry of a paratope.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Multidisciplinary Sciences
Scott A. Jones, Prerana Gogoi, Jonathan J. Ruprecht, Martin S. King, Yang Lee, Thomas Zogg, Els Pardon, Deepak Chand, Stefan Steimle, Danielle M. Copeman, Camila A. Cotrim, Jan Steyaert, Paul G. Crichton, Vera Moiseenkova-Bell, Edmund R. S. Kunji
Summary: Mitochondrial uncoupling protein 1 (UCP1) is responsible for the ability of brown adipose tissue in mammals to burn calories as heat for thermoregulation. UCP1 can be activated by fatty acids to generate heat by catalyzing the leakage of protons across the mitochondrial inner membrane. However, purine nucleotides can bind and inhibit UCP1, regulating proton leak through an unknown molecular mechanism. The cryo-electron microscopy structure of the GTP-inhibited state of UCP1 provides insights into the specific interactions and pH dependency of the regulatory mechanism, indicating that inhibitor binding prevents the conformational changes necessary for proton leak.
Article
Biochemical Research Methods
A. Breine, K. Van Holsbeeck, C. Martin, S. Gonzalez, M. Mannes, E. Pardon, J. Steyaert, H. Remaut, S. Ballet, C. van der Henst
Summary: Membrane interactions greatly influence the mode of action of proteins, cell-penetrating peptides, and antimicrobial peptides. A recent study discovered a nanobody that interacts with the multidrug-resistant bacteria Acinetobacter baumannii, but it only binds to fixed cells. To overcome this limitation, linear peptides corresponding to the complementarity-determining regions (CDR) were synthesized and labeled with fluorescent tags. Microscopy data showed that the CDR3 sequence has clear membrane interactions with living A. baumannii cells, indicating its importance in binding and avoiding cell permeabilization. The cyclization of the peptide with a rigidifying 1,2,3-triazole bridge retained its binding ability and provided proteolytic protection. Overall, this study led to the discovery of novel peptides that bind a multidrug-resistant pathogen.
BIOCONJUGATE CHEMISTRY
(2023)
Article
Multidisciplinary Sciences
Arum Wu, David Salom, John D. Hong, Aleksander Tworak, Kohei Watanabe, Els Pardon, Jan Steyaert, Hideki Kandori, Kota Katayama, Philip D. Kiser, Krzysztof Palczewski
Summary: By discovering specific Nbs that bind to the extracellular surface of rhodopsin and modulate the thermodynamics of its activation process, researchers have revealed the secondary structure of Nbs and how they affect GPCR signaling states. Nbs also improve protein misfolding in disease-associated mutant rhodopsin, making them potential therapeutic agents for related diseases.
NATURE COMMUNICATIONS
(2023)
Meeting Abstract
Biochemistry & Molecular Biology
F. Vascon, L. Maso, M. Chinellato, Y. Bouchiba, E. Campagnaro, S. De Felice, F. Goormaghtigh, P. Bellio, G. Cioci, A. Angelini, G. Celenza, S. Barbe, L. Van Melderen, J. Steyaert, E. Pardon, D. Tondi, L. Cendron