Article
Chemistry, Analytical
Martijn M. Vanduijn, Hendrik-Jan Brouwer, Pablo Sanz de la Torre, Jean-Pierre Chervet, Theo M. Luider
Summary: The research team reported an improved electrochemical method that achieves complete reduction of both intermolecular and intramolecular disulfide bridges in a set of monoclonal antibodies. The method utilizes an online electrochemical flow cell to provide information on the pairing of heavy and light chains in the antibodies. The complete reduction of intramolecular disulfide bridges is important for the intact mass of the antibody chain and the confirmation of the protein's amino acid sequence.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Multidisciplinary
Oliver J. Hale, Helen J. Cooper
Summary: This study demonstrates a mass spectrometry analysis method for the direct analysis of intact oligomeric membrane proteins from tissue. By characterizing protein assembly directly from sheep eye lens sections without sample pre-treatment, our approach allows for simultaneous analysis of soluble protein assemblies.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Multidisciplinary Sciences
Xiaowen Liu, Zhuxin Dong, Gregory Timp
Summary: This article introduces a method of reading protein sequences using sub-nanometer pores and presents computational methods for processing nanospectra and predicting theoretical nanospectra.
SCIENTIFIC REPORTS
(2022)
Article
Chemistry, Analytical
Harvey E. Johnston, Kranthikumar Yadav, Joanna M. Kirkpatrick, George S. Biggs, David Oxley, Holger B. Kramer, Rahul S. Samant
Summary: Complete and reproducible extraction of protein is crucial for proteome analysis. Solvent precipitation SP3 (SP4) is proposed as an alternative method, which captures protein aggregates by centrifugation instead of magnetism. SP4 provides equivalent or higher protein yields and improved reproducibility. It also recovers more low-solubility and transmembrane proteins, offering higher recovery rates.
ANALYTICAL CHEMISTRY
(2022)
Article
Multidisciplinary Sciences
Sarah M. Fantin, Kristine F. Parson, Pramod Yadav, Brock Juliano, Geoffrey C. Li, Charles R. Sanders, Melanie D. Ohi, Brandon T. Ruotolo
Summary: Peripheral myelin protein (PMP22) misfolding is identified as a key factor in various peripheral neuropathies. Mutant forms of PMP22 exhibit differences in stability and propensity to form homodimeric complexes compared to wild-type protein. The formation of PMP22 dimers from destabilized monomers is proposed as a key element in PMP22 mistrafficking, based on combined findings from native ion mobility-mass spectrometry and cellular data.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2021)
Article
Multidisciplinary Sciences
Xiaoran Roger Liu, Yining Huang
Summary: This article reviews several methods for studying protein structure and interactions using mass spectrometry, including hydrogen-deuterium exchange mass spectrometry, hydroxyl radical protein footprinting, and fast photochemical oxidation of proteins.
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
(2023)
Article
Chemistry, Multidisciplinary
Jiyeon Lee, Dahye Im, Yajie Liu, Jing Fang, Xibao Tian, Minsu Kim, Wen-Bin Zhang, Jongcheol Seo
Summary: A technique combining ion mobility spectrometry-mass spectrometry (IMS-MS) and supercharging electrospray ionization (ESI) has been demonstrated to effectively differentiate protein chemical topology. By incorporating as many charges as possible into proteins via supercharging ESI, the protein chains can be largely unfolded and stretched, revealing their hidden chemical topology. Different chemical topologies result in different geometric sizes of the unfolded proteins due to constraints in torsional rotations. The introduction of new topological indices allows for successful differentiation of various protein chemical topologies. This technique provides an easy classification of the topological features of complex proteins, previously concealed by their inherent complexity, and establishes a visual standard for discerning protein topologies.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Multidisciplinary Sciences
Tara K. Bartolec, Xabier Vazquez-Campos, Alexander Norman, Clement Luong, Marcus Johnson, Richard J. Payne, Marc R. Wilkins, Joel P. Mackay, Jason K. K. Low
Summary: Significant advances in cryoelectron microscopy have expanded our ability to create structural models of proteins and protein complexes. Cross-linking mass spectrometry (XL-MS) is demonstrated to be a powerful tool for high-throughput experimental assessment of protein structures and interactions. The combination of XL-MS data with AlphaFold2 predictions offers opportunities to explore the structural proteome and interactome in depth.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2023)
Article
Chemistry, Applied
Xiaoyan Tan, Xinle Tan, Enpeng Li, Yeming Bai, Thoa T. L. Nguyen, Robert G. Gilbert
Summary: The study aimed to explore the relationships between the molecular fine structure of starch and the composition of storage proteins and metabolic enzymes in chickpea seeds. Results showed that storage proteins and starch biosynthetic enzymes influence each other, with a positive correlation between amylopectin size and total protein content. Higher seed globulin content may indicate higher amylose content and more short-medium amylose chains. This research could aid in selecting chickpea varieties with desired qualities, such as low starch digestibility.
CARBOHYDRATE POLYMERS
(2021)
Article
Chemistry, Analytical
Wassim Gabriel, Matthew The, Daniel P. Zolg, Florian P. Bayer, Omar Shouman, Ludwig Lautenbacher, Karsten Schnatbaum, Johannes Zerweck, Tobias Knaute, Bernard Delanghe, Andreas Huhmer, Holger Wenschuh, Ulf Reimer, Guillaume Medard, Bernhard Kuster, Mathias Wilhelm
Summary: The accurate prediction of fragment ion intensities and retention time for peptides has been a topic of significant interest, particularly for unlabeled peptides. In this study, a comprehensive resource of synthetic TMT-labeled peptides was generated as part of the ProteomeTools project, and the deep learning model Prosit was extended to accurately predict the retention time and fragment ion intensities of TMT-labeled peptides with high accuracy.
ANALYTICAL CHEMISTRY
(2022)
Article
Chemistry, Analytical
Lars Kolbowski, Swantje Lenz, Lutz Fischer, Ludwig R. Sinn, Francis J. O'Reilly, Juri Rappsilber
Summary: This study examines the advantages and disadvantages of using MS-cleavable crosslinkers in proteome-wide crosslinking mass spectrometry studies. It is found that the main advantage of MS cleavability of the crosslinker is improved peptide backbone fragmentation, reducing the ambiguity of peptide identifications. Additionally, simpler and faster HCD method outperforms more intricate MS3-based data acquisition approaches.
ANALYTICAL CHEMISTRY
(2022)
Article
Biochemical Research Methods
Shon Cohen, Dina Schneidman-Duhovny
Summary: Large protein complexes cannot be predicted using deep learning, but computational integration of data from experimental techniques can characterize these complexes. Crosslinking mass spectrometry provides spatial information about crosslinked residues. Designing a scoring function to interpret crosslinking data is challenging, and we propose a deep learning model to predict the optimal distance range between crosslinked residue pairs based on their neighborhood structures. Our model achieves area under the receiver-operator curve values of 0.86 and 0.7 for intra- and inter-protein crosslinks, respectively. The deep scoring function has potential applications in structure modeling.
Article
Chemistry, Analytical
Na Wu, Lingyi Jiao, Matthias Butikofer, Zhihui Zeng, Renato Zenobi
Summary: This study successfully utilized commercially available purified proteins as internal standards for high-mass MALDI-MS analysis, achieving absolute quantification of various high-mass analytes. By systematically evaluating four sample deposition methods and using a sandwich deposition method with saturated sinapinic acid, robust quantitative analysis by high-mass MALDI-MS was performed. Furthermore, combining this quantitative strategy with chemical cross-linking allowed for evaluation of the affinity of protein-protein interactions, providing dissociation constants between 10 nM and 5 mu M for the protein complexes formed.
ANALYTICAL CHEMISTRY
(2021)
Article
Plant Sciences
Wiebke Hanke, Julian Patt, Judith Alenfelder, Jan H. Voss, Mitja M. Zdouc, Stefan Kehraus, Jung Bong Kim, Goran Grujicic, Vigneshwaran Namasivayam, Raphael Reher, Christa E. Mueller, Evi Kostenis, Max Cruesemann, Gabriele M. Koenig
Summary: This study investigated and compared the metabolomes of two FR producers using feature-based molecular networking, leading to the discovery of 30 previously unknown FR derivatives, with one-third being unique to C. vaccinii. A novel FR derivative, FR-6, was isolated and confirmed to effectively suppress G(q) signaling. This work demonstrates the power of molecular networking in guiding the discovery of novel G(q)-inhibiting FR derivatives.
JOURNAL OF NATURAL PRODUCTS
(2021)
Article
Chemistry, Analytical
Vilmos Kertesz, Muneeba Khalid, Scott T. Retterer, John F. Cahill
Summary: This study presents a novel strategy to optimize the number and location of sampling sites on microporous membrane-covered microfluidic devices, taking into consideration the channel structure of the device. This strategy reduces the acquisition time and minimizes stress on plants during in situ analysis.
ANALYTICAL CHEMISTRY
(2023)
Article
Biophysics
Hanif M. Khan, Jiqing Guo, Henry J. Duff, D. Peter Tieleman, Sergei Y. Noskov
Summary: Researchers refined the structure of hERG using molecular dynamics flexible fitting (MDFF), recovering missing functionally relevant salt bridges and confirming their functional relevance through electrophysiology experiments. This study demonstrates how refinement of a high-resolution cryoEM structure can bridge the gap between structure and function in the voltage-sensing domain (VSD) of hERG.
BIOPHYSICAL JOURNAL
(2021)
Article
Chemistry, Analytical
Asmita Poudel, George Gachumi, Randy Purves, Ildiko Badea, Anas El-Aneed
Summary: The new LC-MS/MS method allows for quantitative and qualitative analysis of phytosterol oxidation products, providing a rapid and accurate way to analyze the content and structure of 16 POPs in different phytosterols, applicable to various sources of phytosterols. The addition of tocopherols significantly increases the stability of phytosterols in liposomal formulations.
ANALYTICA CHIMICA ACTA
(2022)
Article
Agriculture, Multidisciplinary
Randy W. Purves, Kim Souster, Michelle West, Azhar M. Huda, Caleb M. E. Fisher, Michael W. Belford, Bryn O. Shurmer
Summary: Thyreostatic drugs are illegally used in animals for food production, leading to lower quality meat and potential adverse effects on humans. Detecting these drugs in animal tissues can be challenging due to their low-molecular-weight properties. The study proposes an improved LC-FAIMS-SRM method, which combines liquid chromatography with high-field asymmetric waveform ion mobility spectrometry and selective reaction monitoring. This method demonstrates significant improvements in signal-to-noise ratio and lower limit of quantification compared to the traditional LC-SRM method.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2022)
Article
Multidisciplinary Sciences
Valentina Corradi, Anna N. Bukiya, Williams E. Miranda, Meng Cui, Leigh D. Plant, Diomedes E. Logothetis, D. Peter Tieleman, Sergei Y. Noskov, Avia Rosenhouse-Dantsker
Summary: This study reveals the molecular mechanism of cholesterol's effect on ion channel activity. Through mutagenesis and simulation experiments on the Kir3.4 channel, researchers identified a molecular switch that controls the regulation of the channel by cholesterol. This switch changes the conformation of the channel, leading to different effects of cholesterol on the channel's activity.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Chemistry, Multidisciplinary
Van A. Ngo, Maria Queralt-Martin, Farha Khan, Lucie Bergdoll, Jeff Abramson, Sergey M. Bezrukov, Tatiana K. Rostovtseva, David P. Hoogerheide, Sergei Yu Noskov
Summary: The voltage-dependent anion channel (VDAC) responds to transmembrane potential by gating, and the residue K12 is responsible for its voltage sensitivity.
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
(2022)
Article
Biophysics
Meruyert Kudaibergenova, Jiqing Guo, Hanif M. Khan, James Lees-Miller, Mahdi Mousaei, Williams Miranda, Van A. Ngo, Sergei Yu Noskov, Henry J. Duff
Summary: This study discovered a cationic leak current known as omega current, which arises from mutations of the first charged residue in the S4 of the voltage sensor domains of sodium and potassium voltage-gated channels. By mutating the first charged residue of hERG1, researchers discovered a novel omega current. Molecular dynamics simulations and replica-exchange umbrella sampling simulations revealed the atomistic underpinnings of this mechanism.
BIOPHYSICAL JOURNAL
(2022)
Article
Agronomy
Shaoming Huang, Haixia Zhang, Randy W. W. Purves, Rosalind Bueckert, Bunyamin Tar'an, Thomas D. D. Warkentin
Summary: In this study, the relationship between heat tolerance and the regulation of HSP and ABA hormones was investigated in Canadian pea varieties. The transcription of HSP genes and the accumulation of ABA were upregulated under heat stress. Heat-tolerant varieties showed higher ABA synthesis and turnover rate at 3 hours of heat stress.
Article
Parasitology
Vladislav A. Lobanov, Kelly A. Konecsni, Randy W. Purves, W. Brad Scandrett
Summary: This study developed a recombinant protein of Trichinella spiralis serpin using recombinant technology and applied it in indirect ELISA for detecting exposure to Trichinella in pigs. The Serpin ELISA showed high specificity and sensitivity in commercial pig herds, but lower sensitivity for detecting antibodies induced by experimental infections of pigs with sylvatic Trichinella species.
VETERINARY PARASITOLOGY
(2022)
Article
Chemistry, Applied
Darshika Pathiraja, Janitha P. D. Wanasundara, Fatma M. Elessawy, Randy W. Purves, Albert Vandenberg, Phyllis J. Shand
Summary: This study investigated the water-soluble phenolic compounds and antioxidant activities of lentils with normal-tannin and low-tannin seed coats. The results showed significant variations in total phenolic content among different genotypes, ranging from 1519 to 6502 μg/g. The antioxidant activities were 6-9 times higher in normal-tannin lentils than in low-tannin lentils, indicating the presence of water-extractable natural phenolic compounds with promising antioxidant potential.
Article
Chemistry, Applied
Haixia Zhang, Randy W. Purves, Thomas D. Warkentin, Albert Vandenberg
Summary: Faba bean seeds are a rich source of proteins and levodopa, but the presence of vicine and convicine limits its consumption. A quantification method using FAIMS and QqQ was previously reported, and a similar method with commercial instruments was developed. Both methods provided accurate measurements and recovery, but the commercial method had a lower limit of quantification and a wider linear range. The results showed that levodopa levels in faba bean were genotype dependent, but not directly correlated with vicine or convicine levels.
Article
Agriculture, Multidisciplinary
Fatma M. Elessawy, Derek Wright, Albert Vandenberg, Anas El-Aneed, Randy W. Purves
Summary: Using mass spectrometry-based untargeted metabolomics, this study found that patterned green lentil seed coats contained 28 significantly upregulated polyphenolic metabolites, 19 of which were identified as flavones. Non-patterned green lentil seed coats had almost no flavones, indicating a blockage in their flavone biosynthetic pathway. Although the black pattern is not easily discernible, black seed coats also contained similar flavones as those found in green marbled seed coats, suggesting a marbled pattern on black seed coats.
JOURNAL OF AGRICULTURAL AND FOOD CHEMISTRY
(2023)
Article
Plant Sciences
Stanley Adobor, Sabine Banniza, Albert Vandenberg, Randy W. Purves
Summary: Stemphylium botryosum alters lentil secondary metabolism and has different effects on resistant and susceptible genotypes. Untargeted metabolomics identifies metabolites and biosynthetic pathways that play a crucial role in resistance to S. botryosum. This research contributes to understanding the regulation and reprogramming of lentil metabolism under biotic stress and provides potential targets for breeding enhanced disease resistance.
Article
Endocrinology & Metabolism
Fatma M. M. Elessawy, Jessa Hughes, Hamid Khazaei, Albert Vandenberg, Anas El-Aneed, Randy W. W. Purves
Summary: The study aims to explore the polyphenol diversity and differences in standard and wing petals of faba bean flowers, and identify glycosylated flavonoids that contribute to flower color. It was found that the color variation between the faba bean flowers is mainly controlled by anthocyanins in brown, red, and purple-red flowers, while other flavonoids have different levels of influence in different flower color groups, especially in mixed colors.
Article
Agronomy
Winda Puspitasari, Bjoern Allemann, Deepti Angra, Helen Appleyard, Wolfgang Ecke, Christian Moellers, Tanja Nolte, Randy W. Purves, Carsten Renner, Thomas Robertson-Shersby-Harvie, Rebecca Tacke, Alex Windhorst, Sonja Yaman, Wolfgang Link
Summary: This study applied GWAS to antinutritive compounds in winter faba bean and predicted data for different inbred lines using NIRS technology. It found high heritability for V and V + C, lower for C, and identified several SNPs significantly associated with V and V + C.
JOURNAL FUR KULTURPFLANZEN
(2022)
Article
Biochemical Research Methods
Christopher R. Conant, Isaac K. Attah, Sandilya V. B. Garimella, Gabe Nagy, Aivett Bilbao, Richard D. Smith, Yehia M. Ibrahim
Summary: In this study, five different waveforms were characterized in terms of their transmission efficiency, IMS resolution, and resolving power, with triangle and asymmetric ramp profiles found to offer modestly greater resolution and resolving power compared to others. The research also revealed that separations were generally improved with higher TW amplitudes, moderately improved by lower speeds, but experienced decreases in signal intensity under extreme operating conditions.
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
(2021)