Article
Biotechnology & Applied Microbiology
Hongxu Sun, Hongrui Wang, Qiwei Chen, Wenge Dong, Chao Gao, Haiyan Song, Hui Peng, Ren Li, Hao Wu, Liangyu Hou, Yanhong Chang, Hui Luo
Summary: Protein purification is a fundamental technique in biological research and industrial production, and there is a constant pursuit of efficient, convenient, economical, and environmentally friendly methods. This study discovered that certain cations, including alkaline earth metals, alkali metals, and even nonmetals, can precipitate multi-histidine-tagged proteins at low salts concentrations. Based on this finding, a novel cation affinity purification method was developed that requires only three centrifugal separations and achieves high purity similar to immobilized metal affinity chromatography. The study also provides insights into unexpected protein precipitation and highlights the importance of considering the influence of cations on experimental results. The interaction between histidine-tagged proteins and cations may have broad application prospects.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Natalia Zareba, Lukasz Lewandowski, Dominika Kunachowicz, Rene Kizek, Marta Kepinska
Summary: This study reveals that quantum dots (QDs) can influence the activity of superoxide dismutase (SOD) in a size-dependent manner. Additionally, the migration time of QDs-SOD1 complexes varies depending on the type of QD used, with different effects on fluorescence signals.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Neurosciences
Ian D. Tomlinson, Oleg Kovtun, Ruben Torres, Laurel G. Bellocchio, Travis Josephs, Sandra J. Rosenthal
Summary: The study focused on developing a novel ligand, IDT785, that effectively studied the cellular trafficking and membrane dynamics of SERT, providing valuable insights into SERT localization and function. The ligand demonstrated biological activity by inhibiting SERT-mediated reuptake of serotonin and enabling membrane labeling of SERT for further research.
FRONTIERS IN CELLULAR NEUROSCIENCE
(2021)
Article
Biochemistry & Molecular Biology
Jisoo Park, Sangwoo Lee, Yuseon Kim, Tae Hyeon Yoo
Summary: Antibody-protein conjugates are valuable tools for studying biological systems and advancing therapeutics and diagnostics. Challenges in generating bispecific antibodies and antibody-enzyme conjugates can be addressed through enzymatic and chemical approaches, offering solutions for site-specific conjugation of antibodies with other proteins.
BIOORGANIC & MEDICINAL CHEMISTRY
(2021)
Article
Chemistry, Physical
Zhiyuan Han, Rohit M. Vaidya, Opeyemi H. Arogundade, Liang Ma, Mohammad U. Zahid, Suresh Sarkar, Chia-Wei Kuo, Paul R. Selvin, Andrew M. Smith
Summary: Quantum dots (QDs) are widely used semiconductor nanocrystals in life science studies. A focus of research is to reduce the size of QDs to enhance their accessibility to biological targets. By coating QDs with multidentate polymer coatings, the researchers were able to achieve compact and stable QDs with smaller hydrodynamic sizes. The study also found that the structural characteristics of the polymers play a crucial role in determining the hydrodynamic size, colloidal stability, and biomolecular interactions of the coated QDs. The findings provide insights for the design of next-generation QDs with sizes approaching fluorescent protein labels.
CHEMISTRY OF MATERIALS
(2022)
Article
Biochemical Research Methods
Foo Hou Tan, Jeck Fei Ng, Noorjahan Banu Mohamed Alitheen, Azira Muhamad, Chean Yeah Yong, Khai Wooi Lee
Summary: This study presents a non-chromatographic method for the purification of virus-like particles (VLPs) using C-terminally His-tagged TYMVc (TYMVcHis6) as a protein model. The results show that nickel affinity precipitation can successfully recover over 50% of TYMVcHis6 with a purity above 0.90, and the formed VLPs exhibit similar structures to those purified using conventional chromatography methods.
JOURNAL OF VIROLOGICAL METHODS
(2023)
Article
Physics, Atomic, Molecular & Chemical
Amber A. Flores, Oladapo S. Falokun, Ayobami B. Ilesanmi, Anna V. Arredondo, Linh Truong, Nayeli Fuentes, Riccardo Spezia, Laurence A. Angel
Summary: A comparison was made between a new series of metal binding heptapeptides to evaluate their complex formation with Co(II), Ni(II), and Zn(II) ions. The inclusion of specific amino acid residues at certain positions showed preferences for different metal ions, suggesting potential applications for selective metal binding in biological and analytical processes.
INTERNATIONAL JOURNAL OF MASS SPECTROMETRY
(2021)
Article
Engineering, Electrical & Electronic
Maximilian Bettenhausen, Soenke Gruessing, Elena Hardt, Julia Flesch, Friedhard Roemer, Carlos Alvarado Chavarin, Wolfgang M. Klesse, Changjiang You, Jacob Piehler, Giovanni Capellini, Bernd Witzigmann
JOURNAL OF INFRARED MILLIMETER AND TERAHERTZ WAVES
(2019)
Article
Biochemical Research Methods
Julia Flesch, Marie Kappen, Christoph Drees, Changjiang You, Jacob Piehler
ANALYTICAL AND BIOANALYTICAL CHEMISTRY
(2020)
Article
Biochemistry & Molecular Biology
Hai Pan, Miao Jin, Ashwin Ghadiyaram, Parminder Kaur, Henry E. Miller, Hai Minh Ta, Ming Liu, Yanlin Fan, Chelsea Mahn, Aparna Gorthi, Changjiang You, Jacob Piehler, Robert Riehn, Alexander J. R. Bishop, Yizhi Jane Tao, Hong Wang
NUCLEIC ACIDS RESEARCH
(2020)
Article
Cell & Tissue Engineering
Yi Miao, Andrew Ha, Wim de Lau, Kanako Yuki, Antonio J. M. Santos, Changjiang You, Maarten H. Geurts, Jens Puschhof, Cayetano Pleguezuelos-Manzano, Weng Chuan Peng, Ramazan Senlice, Carol Piani, Jan W. Buikema, Oghenekevwe M. Gbenedio, Mario Vallon, Jenny Yuan, Sanne de Haan, Wieger Hemrika, Kathrin Rosch, Luke T. Dang, David Baker, Melanie Ott, Philippe Depeille, Sean M. Wu, Jarno Drost, Roeland Nusse, Jeroen P. Roose, Jacob Piehler, Sylvia F. Boj, Claudia Y. Janda, Hans Clevers, Calvin J. Kuo, K. Christopher Garcia
Article
Nanoscience & Nanotechnology
Julia Flesch, Maximilian Bettenhausen, Marcin Kazmierczak, Wolfgang M. Klesse, Oliver Skibitzki, Olympia E. Psathaki, Rainer Kurre, Giovanni Capellini, Subhajit Guha, Thomas Schroeder, Bernd Witzigmann, Changjiang You, Jacob Piehler
Summary: Label-free optical detection of biomolecules is currently limited by a lack of specificity rather than sensitivity. The engineered three-dimensional IR-resonant silicon micropillar arrays (Si-MPAs) for protein sensing utilize the characteristic refractive index dispersion in the mid-infrared (IR) regime. The spatially controlled surface functionalization enables efficient targeting of soluble and membrane proteins into sensing hotspots directly from cells cultured on Si-MPAs.
ACS APPLIED MATERIALS & INTERFACES
(2021)
Article
Physics, Applied
Lara Jorde, Zehao Li, Adrian Poeppelwerth, Jacob Piehler, Changjiang You, Carola Meyer
Summary: By designing a surface architecture for reversible protein immobilization on CNTs, we have successfully fabricated reusable biosensors for multiple protein analysis. The specific capturing of His-tagged proteins was achieved by combining Py-PEG-tris-NTA and Ni(II) ions, with the protein binding capacity being restored by imidazole wash.
JOURNAL OF APPLIED PHYSICS
(2021)
Article
Biology
Nadia Fuellbrunn, Zehao Li, Lara Jorde, Christian P. Richter, Rainer Kurre, Lars Langemeyer, Changyuan Yu, Carola Meyer, Jorg Enderlein, Christian Ungermann, Jacob Piehler, Changjiang You
Summary: This study introduces a method called GIET to probe the axial orientation of arrested macromolecules at lipid monolayers, and analyzes the conformational organization of proteins and complexes involved in tethering and fusion at yeast vacuoles. The results reveal that the membrane-anchored Ypt7 protein undergoes conformational reorganization upon interactions with effector proteins, and the tethering complex alternates between 'closed' and 'open' conformations dynamically when recruited to membranes by Ypt7.
Article
Cell Biology
Anthony Fernandez, Markville Bautista, Liying Wu, Fabien Pinaud
Summary: Using single-molecule tracking and super-resolution microscopy, we investigated the nanoscale organization of emerin in the nuclear envelope. We found that emerin forms localized oligomeric nanoclusters stabilized by lamin A/C and the LINC complex. Its interactions with nuclear actin and BAF modulate membrane mobility and oligomerization. In response to mechanical challenges, emerin's mechanotransduction functions and oligomeric state change, leading to nuclear shape adaptation. Abnormal nuclear envelope deformations in EDMD emerin mutants result from improper formation of lamin A/C and LINC complex-stabilized emerin oligomers.
JOURNAL OF CELL SCIENCE
(2022)
Article
Biochemistry & Molecular Biology
Parminder Kaur, Ryan Barnes, Hai Pan, Ariana C. Detwiler, Ming Liu, Chelsea Mahn, Jonathan Hall, Zach Messenger, Changjiang You, Jacob Piehler, Robert C. Smart, Robert Riehn, Patricia L. Opresko, Hong Wang
Summary: The telomere specific shelterin complex including TIN2 plays a crucial role in regulating telomeric DNA structures and interactions between TRF1, TRF2, and TPP1. Knockdown of TIN2 results in the removal of TRF1 and TRF2 from telomeres, affecting downstream effects in cell-based assays.
NUCLEIC ACIDS RESEARCH
(2021)
Article
Multidisciplinary Sciences
Grace W. Chong, Sahand Pirbadian, Yunke Zhao, Lori A. Zacharoff, Fabien Pinaud, Mohamed Y. El-Naggar
Summary: The diffusive dynamics of multiheme cytochromes involved in long-distance electron transfer have not been observed or quantified. In this study, the lateral diffusive dynamics of outer membrane-associated multiheme cytochromes MtrC and OmcA in Shewanella oneidensis were investigated using quantum dot labeling, total internal reflection fluorescence microscopy, and single-particle tracking. Confined diffusion behavior was observed along cell surfaces and membrane nanowires, providing evidence for electron transport via overlap of cytochrome trajectories.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Biochemistry & Molecular Biology
Debika Ojha, Malgorzata M. Jaszczur, Adhirath Sikand, John P. McDonald, Andrew Robinson, Antoine M. van Oijen, Chi H. Mak, Fabien Pinaud, Michael M. Cox, Roger Woodgate, Myron F. Goodman
Summary: Homologs of mutagenic DNA polymerase V are encoded by pathogens and mobile elements. These polymerases can be transferred horizontally into different cells but require activation by host factors.
NUCLEIC ACIDS RESEARCH
(2022)
Article
Biochemical Research Methods
Junel Sotolongo Bellon, Oliver Birkholz, Christian P. Richter, Florian Eull, Hella Kenneweg, Stephan Wilmes, Ulrich Rothbauer, Changjiang You, Mark R. Walter, Rainer Kurre, Jacob Piehler
Summary: Localization and tracking of individual receptors by single-molecule imaging allows for the study of assembly and dynamics of signaling complexes in the plasma membrane. This article presents a comprehensive workflow for imaging and analyzing receptor diffusion and interaction in live cells using up to four colors. The use of engineered GFP variants and fluorescent dyes enables the quantitative resolution of the stoichiometry and dynamics of the IFNg receptor signaling complex in living cells.
CELL REPORTS METHODS
(2022)
Article
Physics, Fluids & Plasmas
Ahis Shrestha, Fabien Pinaud, Christoph A. Haselwandter
Summary: Research has shown that the shape of caveolae and their sensitivity to changes in membrane tension depend strongly on the caveola's spontaneous curvature and the size of caveola domains. Heterogeneity in caveola shape produces a staggered response of caveolae to mechanical perturbations of the cell membrane.
Article
Biology
Chun-Chih Tseng, Ramunas Stanciauskas, Pu Zhang, Dennis Woo, Kaijin Wu, Kevin Kelly, Parkash S. Gill, Min Yu, Fabien Pinaud, Amy S. Lee
LIFE SCIENCE ALLIANCE
(2019)
Article
Chemistry, Multidisciplinary
Michael Philippi, Changjiang You, Christian P. Richter, Mercedes Schmidt, Jannis Thien, Domenik Lisse, Joachim Wollschlaeger, Jacob Piehler, Martin Steinhart
