Article
Biology
Yunfan Fu, Justin Bedo, Anthony T. Papenfuss, Alan F. Rubin
Summary: In this study, a linear regression-based predictor was extended to improve the prediction of amino acid variant impacts by incorporating data from alanine scanning. The results showed that the improvement in model performance is closely related to the correlation between DMS and AS results.
Article
Biochemistry & Molecular Biology
Signe Christensen, Camille Wernersson, Ingemar Andre
Summary: Characterizing the effects of mutations on stability is critical for understanding the function and evolution of proteins and improving their biophysical properties. A multiplex assay for high throughput screening of protein folding was developed, which can identify stabilizing mutations with a low false positive rate. This method can be combined with functional screens to identify mutations that improve both stability and activity.
JOURNAL OF MOLECULAR BIOLOGY
(2023)
Article
Neurosciences
Eder Gambeta, Maria A. Gandini, Ivana A. Souza, Laurent Ferron, Gerald W. Zamponi
Summary: A novel missense mutation in the CACNA1A gene was identified in a patient with trigeminal neuralgia, which alters the gating properties of the Ca(V)2.1 calcium channel. The mutation leads to changes in the voltage-dependence of activation, inactivation, and calcium-dependent inactivation, suggesting a gain of function effect that may contribute to the development of trigeminal neuralgia.
Article
Biology
Jiahui Chen, Daniel R. Woldring, Faqing Huang, Xuefei Huang, Guo-Wei Wei
Summary: High-throughput deep mutational scanning (DMS) experiments have revolutionized various fields such as protein engineering, drug discovery, immunology, cancer biology, and evolutionary biology by providing systematic understanding of protein functions. However, the enormous mutational space associated with proteins exceeds current experimental capabilities, necessitating alternative approaches for DMS. In this study, we propose a topological deep learning (TDL) paradigm that utilizes a new topological data analysis (TDA) technique based on the persistent spectral theory. Our results demonstrate the accuracy and reliability of the TDL-DMS model in predicting binding interface mutations using SARS-CoV-2 datasets. This finding has significant implications for SARS-CoV-2 variant forecasting, antibody design, vaccine development, precision medicine, and protein engineering.
COMPUTERS IN BIOLOGY AND MEDICINE
(2023)
Article
Biochemical Research Methods
Yongcan Chen, Ruyun Hu, Keyi Li, Yating Zhang, Lihao Fu, Jianzhi Zhang, Tong Si
Summary: In this study, deep mutational scanning was used to systematically analyze oxygen-independent flavin mononucleotide-based fluorescent proteins. New empirical data and design rules were obtained, providing insights for engineering protein variants.
ACS SYNTHETIC BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Ekaterina M. Nestorovich, Sergey M. Bezrukov
Summary: Ion channels exhibit gating behavior regulated by transmembrane voltage. This study explores the voltage-induced closure of beta-barrel channels and suggests that it can be seen as reversible protein denaturation rather than functional gating. The research provides insights into the evolutionary aspect of membrane proteins and their development into voltage-gated channels.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Multidisciplinary Sciences
Fenghui Guan, Tianyu Li, Wei Dong, Rui Guo, Hao Chai, Zhiqiu Chen, Zhong Ren, Yang Li, Sheng Ye
Summary: This study reveals the allosteric mechanism of MthK through analyzing its structure and conducting thermodynamic and electrophysiological measurements. The study found that the conformations of the Ca2+-binding sites in MthK alternate between two quaternary states and exhibit significant differences in Ca2+ affinity. An allosteric model of MthK gating mechanism is proposed, in which a cascade of structural events connect the initial Ca2+-binding to the final changes of the ring structure that open the ion-conduction pore. This mechanical model sheds light on the design of allosteric gating and its potential relevance for other ligand-gated ion channels containing the RCK domain.
NATIONAL SCIENCE REVIEW
(2022)
Article
Chemistry, Multidisciplinary
Qi Xiao, Wei-Wei Haoyang, Tao Lin, Zhan-Ting Li, Dan-Wei Zhang, Jun-Li Hou
Summary: A series of peptide-appended bisresorcinarenes were synthesized, which formed tubular conformation induced by intramolecular hydrogen bonds. These derivatives formed unimolecular artificial transmembrane channels in lipid bilayers for selective transport of monovalent cations, and showed reversible ligand-gating behavior in response to alkyl amine and Cu2+.
CHEMICAL COMMUNICATIONS
(2021)
Article
Biochemical Research Methods
Yuki Ogawa, Yohei Katsuyama, Yasuo Ohnishi
Summary: This study applied deep mutational scanning to alter the ligand specificity of the transcriptional regulator XylS and identified the importance of the G71 residue. The results demonstrate the potential of deep mutational scanning in engineering ligand specificity of transcriptional regulators without full structural information.
ACS SYNTHETIC BIOLOGY
(2022)
Article
Biology
Megan Leander, Zhuang Liu, Qiang Cui, Srivatsan Raman
Summary: This study investigates the location and properties of allosteric hotspots in bacterial allosteric transcription factors (aTFs). It is found that hotspots are distributed protein-wide rather than being restricted to pathways connecting allosteric and active sites. The location of hotspots varies across aTFs, but common signatures emerge when comparing hotspots with long-range interactions. Machine learning models reveal that global structural and dynamic properties are better predictors of hotspots than local and physicochemical properties. Furthermore, a model trained on one protein can predict hotspots in a homolog.
Article
Biochemistry & Molecular Biology
Jorge Fernandez-de-Cossio-Diaz, Guido Uguzzoni, Andrea Pagnani
Summary: A computational method is developed to study the relationship between genotype and fitness, trained on sequencing samples from multiple rounds of screening experiments and tested on large-scale mutational scans. The inferred fitness landscape is robust and exhibits high generalization power.
MOLECULAR BIOLOGY AND EVOLUTION
(2021)
Article
Microbiology
Sihan Wang, Tian-Hao Zhang, Menglong Hu, Kejun Tang, Li Sheng, Mengying Hong, Dongdong Chen, Liubo Chen, Yuan Shi, Jun Feng, Jing Qian, Lifeng Sun, Kefeng Ding, Ren Sun, Yushen Du
Summary: This study provides a comprehensive characterization of the fitness and drug resistance profiles of NA mutants in an in vivo context. It identifies novel drug resistance mutations and a potential allosteric mutation that confers resistance to multiple drugs. The fitness cost associated with drug-resistant mutations may limit their spread.
Article
Biology
Shuo Han, Sophia Peng, Joshua Vance, Kimberly Tran, Nhu Do, Nauy Bui, Zhenhua Gui, Shizhen Wang
Summary: This article investigates the conformational dynamics and gating mechanisms of voltage-gated proton (Hv) channels. By using single-molecule fluorescence resonance energy transfer (smFRET) with purified human voltage-gated proton (hHv1) channels reconstituted in liposomes, the real-time conformational trajectories of the S4 segment were observed. The results demonstrate that voltage and pH gradient can shift the conformational dynamics of the S4 segment, controlling the gating of the channel. The authors propose a kinetic model to explain the voltage and pH gating mechanisms in Hv channels, which may have implications for understanding the voltage sensing and gating in other voltage-gated ion channels.
Review
Virology
Thomas D. Burton, Nicholas S. Eyre
Summary: Recent high-throughput techniques in molecular virology, such as proteomics, CRISPR screens, ChIP-seq experiments, and deep mutational scanning, have revolutionized the field by providing new insights and tools for studying viruses. These techniques have been applied to RNA viruses, including members of the Flaviviridae family, Influenza A Virus, and Severe Acute Respiratory Syndrome Coronavirus 2, with implications for viral replication cycles and potential drug targets.
Article
Virology
Yuan Li, Sarah Arcos, Kimberly R. Sabsay, Aartjan J. W. te Velthuis, Adam S. Lauring
Summary: The influenza virus polymerase plays a central role in the evolution of the virus. Mutations in the polymerase are necessary for the efficient spread of animal-derived viruses in human populations. The polymerase also determines the rate at which the virus acquires mutations that lead to drug resistance or antigenic drift. However, our understanding of the effects of mutations on the influenza RNA-dependent RNA polymerase (RdRp) is limited. In this study, deep mutational scanning of a specific protein in the polymerase was performed to understand the constraints and effects of mutations. The results provide valuable insights into the replication and evolution of the influenza virus.
JOURNAL OF VIROLOGY
(2023)
Article
Multidisciplinary Sciences
Katherine R. Heal, Wei Qin, Francois Ribalet, Anthony D. Bertagnolli, Willow Coyote-Maestas, Laura R. Hmelo, James W. Moffett, Allan H. Devol, E. Virginia Armbrust, David A. Stahl, Anitra E. Ingalls
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2017)
Article
Multidisciplinary Sciences
Willow Coyote-Maestas, Yungui He, Chad L. Myers, Daniel Schmidt
NATURE COMMUNICATIONS
(2019)
Article
Biochemistry & Molecular Biology
Willow Coyote-Maestas, David Nedrud, Steffan Okorafor, Yungui He, Daniel Schmidt
NUCLEIC ACIDS RESEARCH
(2020)
Article
Biochemical Research Methods
Alina C. Zdechlik, Yungui He, Eric J. Aird, Wendy R. Gordon, Daniel Schmidt
BIOCONJUGATE CHEMISTRY
(2020)
Article
Biochemistry & Molecular Biology
David Nedrud, Willow Coyote-Maestas, Daniel Schmidt
Summary: The programmable DMS technique SPINE was used to study epistasis in the PSD95 PDZ3 domain, revealing widespread proximal negative epistasis and strong long-range positive epistasis. This interaction between position pairs facilitated the evolutionary expansion and specialization of PDZ domains, providing insights into protein function and adaptation.
PROTEINS-STRUCTURE FUNCTION AND BIOINFORMATICS
(2021)
Editorial Material
Biology
Willow Coyote-Maestas, James S. Fraser
Summary: A new method of altering the genome of bacteriophages allows for the production of large libraries of variants, enabling the design of these viruses to target species harmful to human health.
Article
Multidisciplinary Sciences
Willow Coyote-Maestas, David Nedrud, Antonio Suma, Yungui He, Kenneth A. Matreyek, Douglas M. Fowler, Vincenzo Carnevale, Chad L. Myers, Daniel Schmidt
Summary: The research conducted a large-scale, high-throughput biochemical assay to determine the compatibility of over 300,000 domain recombination variants of the inward rectifier K+ channel Kir2.1. They found that genomic and biophysical mechanisms acted together to favor gain of large, structured domains at protein termini during ion channel evolution.
NATURE COMMUNICATIONS
(2021)
Article
Biotechnology & Applied Microbiology
Zion Lee, Min Lu, Eesha Irfanullah, Morgan Soukup, Daniel Schmidt, Wei-Shou Hu
Summary: This study describes a stable cell line that can be used to quantify the titers of infectious recombinant adeno-associated viruses (rAAVs). It has high sensitivity and eliminates the need for a helper virus, improving assay consistency and allowing high-throughput applications. This cell line can serve as an effective tool for quantifying infectious rAAV titers to advance gene therapy vector biomanufacturing.
HUMAN GENE THERAPY
(2023)
Article
Biotechnology & Applied Microbiology
Christian B. Macdonald, David Nedrud, Patrick Rockefeller Grimes, Donovan Trinidad, James S. Fraser, Willow Coyote-Maestas
Summary: Insertions and deletions (indels) play a crucial role in evolution and disease. However, their study is often overlooked in mutational scans due to technical challenges. In this research, we developed a cost-effective and unbiased method called DIMPLE to systematically create indels in genes, and applied it to study the impact of indels on potassium channel structure, disease, and evolution. Our findings show that deletions are the most disruptive, beta sheets are highly sensitive to indels, and flexible loops can tolerate insertions but are sensitive to deletions.