Protein Chemistry Research Articles

Protein click chemistry and its potential for medical applications.

A revolution in chemical biology occurred with the introduction of click chemistry. Click chemistry plays an important role in protein chemistry modifications, providing specific, sensitive, rapid, and easy-to-handle methods. Under physiological conditions, click chemistry often overlaps with bioorthogonal chemistry, defined as reactions that occur rapidly and selectively without interfering with biological processes. Click chemistry is used for the posttranslational modification of proteins based on covalent bond formations. With the contribution of click reactions, selective modification of proteins would be developed, representing an alternative to other technologies in preparing new proteins or enzymes for studying specific protein functions in different biological processes. Click-modified proteins have potential in diverse applications such as imaging, labeling, sensing, drug design, and enzyme technology. Due to the promising role of proteins in disease diagnosis and therapy, this review aims to highlight the growing applications of click strategies in protein chemistry over the last two decades, with a special emphasis on medicinal applications.

First person – Emily D. McParland and T. Amber Butcher

ABSTRACT First Person is a series of interviews with the first authors of a selection of papers published in Journal of Cell Science, helping researchers promote themselves alongside their papers. Emily D. McParland and T. Amber Butcher are co-first authors on ‘ The Dilute domain in Canoe is not essential for linking cell junctions to the cytoskeleton but supports morphogenesis robustness’, published in JCS. Emily and Amber conducted the research described in this article while they were postbaccalaureate research assistants in the labs of Dr Mark Peifer (E.D.M. and T.A.B.) and Dr Kevin Slep (T.A.B.) at Chapel Hill Department of Biology, University of North Carolina, USA. Emily is now a PhD student in the lab of Dr Thomas Roberts at Brown University, Providence, USA, where she is broadly interested in functional morphology and muscle biomechanics, and focuses on the function of the collagenous extracellular matrix in muscle force production. Amber is a PhD student in the lab of Dr Eric Greene at Columbia University Department of Biochemistry and Molecular Biophysics, New York, USA, where she uses biochemical and biophysical techniques to investigate protein function, most recently within the context of homologous recombination.

Identification and Characterization of the Gene Responsible for the O3 Mating Type Substance in Paramecium caudatum.

The process of sexual reproduction in eukaryotes starts when gametes from two different sexes encounter each other. Paramecium, a unicellular eukaryote, undergoes conjugation and uses a gametic nucleus to enter the sexual reproductive process. The molecules responsible for recognizing mating partners, hypothetically called mating-type substances, are still unclear. We have identified an O3-type mating substance polypeptide and its gene sequence using protein chemistry, molecular genetics, immunofluorescence, RNA interference, and microinjection. The O3-type substance is a polypeptide found in the ciliary membranes, located from the head to the ventral side of cells. The O3-type substance has a kinase-like domain in its N-terminal part located outside the cell and four EF-hand motifs that bind calcium ions in its C-terminal part located inside the cell. RNA interference and immunofluorescence revealed that this polypeptide positively correlated with the expression of mating reactivity. Microinjection of an expression vector incorporating the O3Pc-MSP gene (Oms3) induced additional O3 mating type in the recipient clones of different mating types or syngen. Phylogenetic analysis indicates that this gene is widely present in eukaryotes and exhibits high homology among closely related species. The O3Pc-MSP (Oms3) gene had nine silent mutations compared to the complementary mating type of the E3 homologue gene.

Hexafluoroisopropanol as a Bioconjugation Medium of Ultrafast, Tryptophan-Selective Catalysis.

The past decade has seen a remarkable growth in the number of bioconjugation techniques in chemistry, biology, material science, and biomedical fields. A core design element in bioconjugation technology is a chemical reaction that can form a covalent bond between the protein of interest and the labeling reagent. Achieving chemoselective protein bioconjugation in aqueous media is challenging, especially for generally less reactive amino acid residues, such as tryptophan. We present here the development of tryptophan-selective bioconjugation methods through ultrafast Lewis acid-catalyzed reactions in hexafluoroisopropanol (HFIP). Structure-reactivity relationship studies have revealed a combination of thiophene and ethanol moieties to give a suitable labeling reagent for this bioconjugation process, which enables modification of peptides and proteins in an extremely rapid reaction unencumbered by noticeable side reactions. The capability of the labeling method also facilitated radiofluorination application as well as antibody functionalization. Enhancement of an α-helix by HFIP leads to its compatibility with a certain protein, and this report also demonstrates a further stabilization strategy achieved by the addition of an ionic liquid to the HFIP medium. The nonaqueous bioconjugation approaches allow access to numerous chemical reactions that are unavailable in traditional aqueous processes and will further advance the chemistry of proteins.

Effects of storage conditions on chemistry and technological properties of different cultivars of Chickpea

The impact of storage conditions, such as 40 °C, high humidities (55% and 65%), and storage (360 days), on the nutritional value, pasting, and functional properties, color differences, and quality of protein and starch in five Chickpea (Cicer arietinum) cultivars (Crown, Royal, Sierra, Orion, and Frontier) was determined. The Sierra cultivar had the highest initial moisture content (MC, 7.7 ± 0.01%), and MC increased over time for all samples stored at 55% and 65% RH and 21 °C. Protein (PC), total starch (TSC), and fat (FC) contents changed in all cultivars during storage. Under the same storage conditions (65% RH and 40 °C treatment), the Frontier variety had significantly higher PC at day zero (24.0 ± 0% dwb) and 360 (23.9 ± 0.2% dwb) compared to other cultivars, while no significant differences existed between other cultivars. A general upward trend in the pasting data was observed for all cultivars of the 360-day stored samples. In contrast, the gel firmness of the gels formed during Rapid Visco Analysis (RVA) was lower for the 360-day samples. Emulsion capacity (EC) and Foaming capacity (FC) changed significantly in all samples over time under the effects of different variables. Color analysis revealed reduced yellowness in all samples during storage. Also, lightness values decreased over time, indicating seed darkening during storage. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed the disappearance of major protein bands around 37 and 55 kDa after 360 days, indicating protein aggregation and structural alterations. Fourier transform infrared spectroscopy (FTIR) of amide II structures indicated interactions and differences in the secondary structure of proteins in the samples stored for 360 days. Starch analysis via SEM revealed protein-coated starch granules, indicating protein-starch interactions occurred during storage. Harsh storage conditions (65% RH and 40 °C) significantly affected nutritional quality, functional and pasting properties, and protein chemistry of chickpeas and should be avoided for extended storage.

Impact of Immunopathy and Coagulopathy on Multi-Organ Failure and Mortality in a Lethal Porcine Model of Controlled and Uncontrolled Hemorrhage.

Uncontrolled hemorrhage is a major preventable cause of death in patients with trauma. However, the majority of large animal models of hemorrhage have utilized controlled hemorrhage rather than uncontrolled hemorrhage to investigate the impact of immunopathy and coagulopathy on multi-organ failure (MOF) and mortality. This study evaluates these alterations in a severe porcine controlled and uncontrolled hemorrhagic shock (HS) model. Anesthetized female swine underwent controlled hemorrhage and uncontrolled hemorrhage by partial splenic resection followed with or without lactated Ringer solution (LR) or Voluven® resuscitation. Swine were surveyed 6 h after completion of splenic hemorrhage or until death. Blood chemistry, physiologic variables, systemic and tissue levels of complement proteins and cytokines, coagulation parameters, organ function, and damage were recorded and assessed. HS resulted in systemic and local complement activation, cytokine release, hypocoagulopathy, metabolic acidosis, MOF, and no animal survival. Resuscitation with LR and Voluven® after HS improved hemodynamic parameters (MAP and SI), metabolic acidosis, hyperkalemia, and survival but resulted in increased complement activation and worse coagulopathy. Compared with the LR group, the animals with hemorrhagic shock treated with Voluven® had worse dilutional anemia, coagulopathy, renal and hepatic dysfunction, increased myocardial complement activation and renal damage, and decreased survival rate. Hemorrhagic shock triggers early immunopathy and coagulopathy and appears associated with MOF and death. This study indicates that immunopathy and coagulopathy are therapeutic targets that may be addressed with a high-impact adjunctive treatment to conventional resuscitation.

Developing peptide-porphyrin conjugates as broad spectrum antivirals, from molecular biophysics to in vivo studies

Developing peptide-porphyrin conjugates as broad spectrum antivirals, from molecular biophysics to in vivo studies

Dual-wavelength metalens enables Epi-fluorescence detection from single molecules

Single molecule fluorescence spectroscopy is at the heart of molecular biophysics research and the most sensitive biosensing assays. The growing demand for precision medicine and environmental monitoring requires the creation of miniaturized and portable sensing platforms. However, the need for highly sophisticated objective lenses has precluded the development of single molecule detection systems for truly portable devices. Here, we propose a dielectric metalens device of submicrometer thickness to excite and collect light from fluorescent molecules instead of an objective lens. The high numerical aperture, high focusing efficiency, and dual-wavelength operation of the metalens enable the implementation of fluorescence correlation spectroscopy with a single Alexa 647 molecule in the focal volume. Moreover, the metalens enables real-time monitoring of individual fluorescent nanoparticle transitions and identification of hydrodynamic diameters ranging from a few to hundreds of nanometers. This advancement in sensitivity extends the application of the metalens technology to ultracompact single-molecule sensors.

Solitary waves of the generalized Zakharov equations via integration algorithms

<abstract><p>In many applications, the investigation of traveling wave solutions is essential in obtaining an accurate description of the dynamical behavior of most physical phenomena. The exact solutions to nonlinear equations can provide more physical descriptions and insightful details for many problems of practical interest. This paper focuses on investigating the solitary wave solutions of the generalized Zakharov equations (GZEs) by using four integration algorithms, namely, the modified $ (g'/g^{2}) $-expansion method, the modified $ (g') $-expansion method, the generalized simple ($ w/g $)-expansion method, and the addendum to Kudryashov's method. The GZEs have been widely used to describe the propagation of Langmuir waves in the field of plasma physics. The efficiency and simplicity of these methods are evaluated based on their application to GZEs, which have yielded multiple new optical solitary wave solutions in the form of rational, trigonometric, and hyperbolic functions. By using a suitable wave transformation, the coupled nonlinear partial differential equations are converted into ordinary differential equations. The derived optical solutions are graphically depicted in $ 2 $D and $ 3 $D plots for some specific parameter values. The traveling wave solutions discovered in the current study constitute just one example of the desired solutions that may enable the exploration of the physical properties of many complex systems and could also contribute greatly to improving our understanding of many interesting natural phenomena that arise in different applications, including plasma physics, fluid mechanics, protein chemistry, wave propagation, and optical fibers.</p></abstract>

Electrochemistry-enabled residue-specific modification of peptides and proteins.

Selective chemical reactions at precise amino acid residues of peptides and proteins have become an exploding field of research in the last few decades. With the emerging utility of bioconjugated peptides and proteins as drug leads and therapeutic agents, the design of smart protocols to modulate and conjugate biomolecules has become necessary. During this modification, the most important concern of biochemists is to keep intact the structural integrity of the biomolecules. Hence, a soft and selective biocompatible reaction environment is necessary. Electrochemistry, a mild and elegant tunable reaction platform to synthesize complex molecules while avoiding harsh and toxic chemicals, can provide such a reaction condition. However, this strategy is yet to be fully exploited in the field of selective modification of polypeptides. With this possibility, the use of electrochemistry as a reaction toolbox in peptide and protein chemistry is flourishing day by day. Unfortunately, there is no suitable review article summarizing the residue-specific modification of biomolecules. The present review provides a comprehensive summary of the latest manifested electrochemical approaches for the modulation of five redox-active amino acid residues, namely cysteine, tyrosine, tryptophan, histidine and methionine, found in peptides and proteins. The article also highlights the incredible potential of electrochemistry for the regio- as well as chemoselective bioconjugation strategy of biomolecules.

Ability of three dairy feed evaluation systems to predict postruminal outflows of amino acids in dairy cows: A meta-analysis

Adequate prediction of postruminal outflows of essential AA (EAA) is the starting point of balancing rations for EAA in dairy cows. The objective of this meta-analysis was to compare the performance of 3 dairy feed evaluation systems (National Research Council [NRC], Cornell Net Protein and Carbohydrate System version 6.5.5 [CNCPS], and National Academies of Sciences, Engineering and Medicine [NASEM]) to predict EAA outflows (Trp was not tested). The data set included a total of 354 treatment means from 70 duodenal and 24 omasal studies. To avoid Type I error, mean and linear biases were considered of concern if statistically significant and representing > 5.0% of the observed mean. Analyses were conducted on raw observed values and on observations adjusted for the random effect of study. The analysis on raw data indicates the ability of the feed evaluation system to predict absolute values whereas the analysis on adjusted values indicates its ability to predict responses of EAA outflows to dietary changes. For the prediction of absolute values (based on raw data), NRC underpredicted outflows of all EAA, from 5.3 to 8.6% of the observed mean (%obs.mean) except for Leu, Lys, and Val; NASEM overpredicted Lys (10.8%obs.mean); and CNCPS overpredicted Arg, His, Lys, Met, and Val (5.2 to 26.0%obs.mean). No EAA had a linear bias of concern with NASEM, followed by NRC for His (6.8%obs.mean), and CNCPS for all EAA (5.6 to 12.2%obs.mean) except Leu, Phe, and Thr. On the other hand, for the prediction of responses to dietary changes (based on adjusted data), NRC had 2 EAA presenting a linear bias of concern, followed by NASEM and CNCPS with 4 and 6 EAA, respectively. Predictions of His showed a linear bias of concern (5.3 to 9.6%obs.mean) with the 3 feed evaluation systems. Measured chemistry of crude protein and EAA were reported for 1 or more feed ingredients of the ration in 36% of the studies, and resulted in decreased linear biases in the 3 feed evaluation systems. The difference in mean biases of Met outflows was systematically positive when comparing omasal versus duodenal studies. Predictions of Met outflows with NRC had a higher concordance correlation coefficient in duodenal (used to develop NRC equations) versus omasal studies, whereas the opposite was observed with CNCPS, the latter showing the lowest mean bias for Met in omasal sampling studies. The 30% difference in Met mean biases between sampling sites appeared related to a similar difference found for observed Met versus NAN outflows between duodenal and omasal studies, which is independent of predictions. In conclusion, NRC and NASEM yielded accurate predictions of EAA outflows, with a small superiority of NASEM to predict absolute values, and slight superiority of NRC to predict the responses to dietary changes. In comparison, CNCPS may present mean and linear biases of concern for many EAA. Moreover, it remains to determine which sampling site is more representative of the true supply of EAA to the cows.

Advances in Microsphere-based Super-resolution Imaging.

Techniques to resolve images beyond the diffraction limit of light with a large field of view (FOV) are necessary to foster progress in various fields such as cell and molecular biology, biophysics, and nanotechnology, where nanoscale resolution is crucial for understanding the intricate details of large-scale molecular interactions. Although several means of achieving super-resolutions exist, they are often hindered by factors such as high costs, significant complexity, lengthy processing times, and the classical tradeoff between image resolution and FOV. Microsphere-based super-resolution imaging has emerged as a promising approach to address these limitations. In this review, we delve into the theoretical underpinnings of microsphere-based imaging and the associated photonic nanojet. This is followed by a comprehensive exploration of various microsphere-based imaging techniques, encompassing static imaging, mechanical scanning, optical scanning, and acoustofluidic scanning methodologies. This review concludes with a forward-looking perspective on the potential applications and future scientific directions of this innovative technology.

Defining Hereditary Pyropoikilocytosis (HPP) in the Molecular Diagnostic Era

Misconceptions continue in distinguishing hereditary elliptocytosis (HE) with hemolysis in early perinatal period and hereditary pyropoikilocytosis (HPP). In 1975, Zarkowsky and colleagues described a group of children with transfusion dependent congenital hemolytic anemia with the unique morphology of “microspherocytes, measuring 2-3 microns in diameter, and cells with blunted projections or triangular in shape”, a morphology resembling the heat induced damage to normal erythrocytes - hence the name hereditary pyropoikilocytosis (HPP). Subsequently, similar cases were described with or without demonstrable heat sensitivity (Ravindranath & Johnson, 1985). Regardless, the common denominators were 1) transfusion dependency early in infancy - resolved with splenectomy, 2) severe microcytosis, often <60 fL, and 3) a decrease in membrane bound spectrin (decreased spectrin/band3 ratio). Family studies showed normal RBC morphology in both parents (Ravindranath & Johnson, 1985) or in some cases, one parent with HE (Zarkowsky et al., 1975). Protein chemistry studies showed decreased tetramer formation and tryptic digests showed mutations within the n-terminal alpha 1 domain of alpha spectrin - the dimer/dimer interaction site. Thus, homozygosity or double heterozygosity of mutations in the dimer/dimer interaction site of alpha spectrin (alpha 1 domain of alpha spectrin) defined HPP while classical HE cases showed heterozygous mutations in the alpha 1 domain. A fact unexplained was how this results in spectrin deficiency, specifically, alpha spectrin deficiency (Ravindranath & Johnson, 1985). Using the molecular analytical tool AnemiaID (courtesy of Agios Pharmaceuticals, Inc.), we show that HPP is defined by the co-inheritance of two SPTA1 mutations and Alpha-LeLy polymorphism ( αLeLy rs28525570; SPTA1[c.5572C>G;c.6531-12C>T]) in one or both alleles, thus explaining the increased propensity for red cell fragmentation and the spectrin deficiency noted on acrylamide gels [Table: Clinical and molecular characteristics of our cases]. Of note, homozygosity for αLeLy polymorphism and one SPTA1 variant did not result in clinical HPP phenotype [HE-2 in Table]. At a clinical laboratory level, the extreme microcytosis in HPP is evident in the markedly diminished band3 content on eosin-5'-maleimide (EMA) binding test- mean channel fluorescence (MCF) of <275.0 (control: 473.0-549.0) while HE (including neonatal HE with hemolysis) show a double peak pattern in EMA (the minor peak representing the fragmented cells while the major peak had MCF higher than control MCF) and the characteristic trapezoid pattern on osmotic gradient ektacytometry (not shown).

Concentrated ionic liquids for proteomics: Caveat emptor!

The use of concentrated ionic liquids (ILs) in the bioanalytical chemistry of proteins is sparse; typically, dilute aqueous IL solutions are used. Concentrated ILs have unique properties that may allow researchers to dissolve previously insoluble protein analytes, to increase the depth and robustness of sample preparation and the analysis of proteins. Previous research using concentrated ILs for this purpose is sparse and there is a need to systematically investigate the structure-activity relationship between the IL structure and its capacity to solubilise proteins. Here, bovine serum albumin was dissolved in various ionic liquids and monitored over time by light microscopy and SDS-PAGE. While qualitative, these measures provide a good estimate of, respectively, the dissolving power of an IL towards the given protein and the retained integrity of the protein. Hydrophilic ILs show the best solubilisation capacity and higher temperatures (in a restricted sense) improve the solubility of the protein. Higher temperatures and longer reaction times reduce the molecular weight of the protein, which could inhibit their applicability in proteomics, unless the conditions are judiciously controlled. Researchers should exercise caution when using concentrated ILs for protein analysis until the full scope and limitations are known, an aspect we are presently investigating.

Chemical Synthesis of Microtubule-Associated Protein Tau.

Deposits of the microtubule-associated protein Tau (MAPT) serve as a hallmark of neurodegenerative diseases known as tauopathies. Numerous studies have demonstrated that in diseases such as Alzheimer's disease (AD), Tau undergoes extensive remodeling. The attachment of post-translational modifications distributed throughout the entire sequence of the protein correlates with clinical presentation. A systematic examination of these protein alterations can shed light on their roles in both healthy and diseased states. However, the ability to access these modifications in the entire protein chain is limited as Tau can only be produced recombinantly or through semisynthesis. In this article, we describe the first chemical synthesis of the longest 2N4R isoform of Tau, consisting of 441 amino acids. The 2N4R Tau was divided into 3 major segments and a total of 11 fragments, all of which were prepared via solid-phase peptide synthesis. The successful chemical strategy has relied on the strategic use of two cysteine sites (C291 and C322) for the native chemical ligations (NCLs). This was combined with modern preparative protein chemistries, such as mercaptothreonine ligation (T205), diselenide-selenoester ligation (D358), and mutations of mercaptoamino acids into native residues via homogeneous radical desulfurization (A40, A77, A119, A157, A246, and A390). The successful completion of the synthesis has established a robust and scalable route to the native protein in multimilligram quantities and high purity. In broader terms, the presented strategy can be applied to the preparation of other shorter isoforms of Tau as well as to introduce all post-translational modifications that are characteristic of tauopathies such as AD.

Meldrum's Acid Furfural Conjugate MAFC: A New Entry as Chromogenic Sensor for Specific Amine Identification.

Bioactive amines are highly relevant for clinical and industrial application to ensure the metabolic status of a biological process. Apart from this, generally, amine identification is a key step in various bioorganic processes ranging from protein chemistry to biomaterial fabrication. However, many amines have a negative impact on the environment and the excess intake of amines can have tremendous adverse health effects. Thus, easy, fast, sensitive, and reliable sensing methods for amine identification are strongly searched for. In the past few years, Meldrum's acid furfural conjugate (MAFC) has been extensively explored as a starting material for the synthesis of photoswitchable donor-acceptor Stenhouse adducts (DASA). DASA formation hereby results from the rapid reaction of MAFC with primary and secondary amines, which has so far been demonstrated through numerous publications for different applications. The linear form of the MAFC-based DASA exhibits intense pink coloration due to its linear conjugated triene-2-ol conformation, which has inspired researchers to use this easy synthesizable molecule as an optical sensor for primary, secondary, and biogenic amines. Due to its new entry into amine identification, a collection of the literature exclusively on MAFC is demanded. In this mini review, we intend to present the state-of-the-art of MAFC as an optical molecular sensor in hopes to motivate researchers to find even more applications of MAFC-based sensors and methods that pave the way to their usage in medicinal applications.

The emerging importance of NAD+ metabolome for nutrition and food sciences: A bibliometric analysis

NAD+ is crucial for cellular balance in all organisms. Recent interest has focused on its metabolism. Deficiency-related issues can be countered using NMN and NR, potential therapies available through diet or supplementation. These compounds are emerging as nutraceuticals and bioactive food elements. Bibliometric analysis was used to study the scientific output and impact of research on the NAD+ metabolome. For this, the terms “nicotinamide adenine dinucleotide” or “NAD+” or “NAD+ precursors” or “nicotinamide riboside” or “nicotinamide mononucleotide” were searched in the topic title in The Web of Science™ database. Only original articles and articles review were considered. VOSviewer was used for constructing and visualizing the bibliometric networks maps. A total of 10,568 papers were published, with a consistent increase in the number of publications, especially over the last decade. Biochemistry molecular biology, biophysics, cell biology, chemistry multidisciplinary and microbiology were the top 5 field research in the subject. The majority of the publications came from the ‘Journal of biological chemistry’ and ‘Biochemistry’ journals. The prominent research topics included longevity, sirtuins, NAD+ precursors, oxidation and reduction reactions, enzymes, microbial biotechnology, cancer therapy, and crop science. Finally, a guide for future investigation in nutrition and food sciences areas was proposed, which holds immense potential for developing innovative strategies to enhance food quality, promote optimal nutrition, and explore new avenues for improving human health. To our knowledge, this is the first bibliometric analysis of NAD+ metabolome, which can provide valuable insights into the research landscape in this multidisciplinary rapidly growing field.

Resonance Raman spectra of blue copper proteins: Variable temperature spectra of Thermus thermophilus HB27 laccase

The resonance Raman (rR) spectra of the oxidized type 1 copper active site (CuT1) in Thermus thermophilus HB27 laccase (Tth-lac) has been determined in the 20 to 80 °C temperature range using 633-nm excitation. The positions and relative intensities of rR peaks are virtually independent of temperature, indicating that CuT1 ligation is robust over the investigated range. The intensity-weighted average of Tth-lac Cu-SCys vibrations (<ν(Cu-SCys)>) = 423 cm−1) is higher than those of most cupredoxins but is comparable to those of other multicopper oxidases (MCOs). <ν(Cu-SCys)> values for Tth-lac and several CuT1 centers in cupredoxins and MCOs do not correlate well with Cu-SCys bond lengths but do exhibit systematic trends with redox thermodynamic properties. PrologueF. Ann Walker was a great scholar and dear friend. While at Columbia in the early 1960s, I (HBG) followed her graduate work at Brown on the effects of axial ligands on vanadyl ion EPR spectra. Dick Carlin, her thesis adviser, invited me to serve as external member of her thesis committee. I joined, made my way to Providence, met her just before the exam, and greatly admired (enjoyed!) her thoughtful responses to questions from physical chemists about metal-oxo electronic structures. Our friendship grew stronger over the years, enhanced by lively discussions of heme protein chemistry in San Francisco, Pasadena, Tucson, and at Gordon Research Conferences. Ann was a superstar in biological inorganic chemistry. She will be sorely missed but not forgotten.

VII congress of biophysicists of Russia

The problems and results of research in biophysics, which were devoted to the VII Congress of Biophysicists of Russia (Krasnodar, April 17-23, 2023, http://rusbiophysics.ru/db/conf.pl), are discussed. The results of fundamental and applied research in the field of molecular biophysics, cell biophysics, biophysics of complex multicomponent systems were presented at plenary, sectional and poster sessions. The structure and dynamics of biopolymers, the fundamental mechanisms underlying the impact of physicochemical factors on biological systems, membrane and transport processes were actively discussed. Much attention was paid to new experimental methods of biophysical research, methods of bioinformatics, computer and mathematical modeling as essential research tools at all levels of organization of living systems. Along with the fundamental problems of studying the biophysical mechanisms of regulation of processes at the molecular, subcellular and cellular levels, much attention was paid to applied research in the field of biotechnology and environmental monitoring. Works in the field of medical biophysics were especially widely represented. During the Congress, the National Council for Biophysics was formed.

Biomolecular NMR in the AI-assisted structural biology era: Old tricks and new opportunities

Over the last 40 years nuclear magnetic resonance (NMR) spectroscopy has established itself as one of the most versatile techniques for the characterization of biomolecules, especially proteins. Given the molecular size limitations of NMR together with recent advances in cryo-electron microscopy and artificial intelligence-assisted protein structure prediction, the bright future of NMR in structural biology has been put into question. In this mini review we argue the contrary. We discuss the unique opportunities solution NMR offers to the protein chemist that distinguish it from all other experimental or computational methods, and how it can benefit from machine learning.