Amino acid selective unlabeling for sequence specific resonance assignments in proteins

Sequence specific resonance assignment constitutes an important step towards high-resolution structure determination of proteins by NMR and is aided by selective identification and assignment of amino acid types. The traditional approach to selective labeling yields only the chemical shifts of the particular amino acid being selected and does not help in establishing a link between adjacent residues along the polypeptide chain, which is important for sequential assignments. An alternative approach is the method of amino acid selective `unlabeling' or reverse labeling, which involves selective unlabeling of specific amino acid types against a uniformly C-13/N-15 labeled background. Based on this method, we present a novel approach for sequential assignments in proteins. The method involves a new NMR experiment named, {(CO)-C-12 (i) -N-15 (i+1)}-filtered HSQC, which aids in linking the H-1(N)/N-15 resonances of the selectively unlabeled residue, i, and its C-terminal neighbor, i + 1, in HN-detected double and triple resonance spectra. This leads to the assignment of a tri-peptide segment from the knowledge of the amino acid types of residues: i - 1, i and i + 1, thereby speeding up the sequential assignment process. The method has the advantage of being relatively inexpensive, applicable to H-2 labeled protein and can be coupled with cell-free synthesis and/or automated assignment approaches. A detailed survey involving unlabeling of different amino acid types individually or in pairs reveals that the proposed approach is also robust to misincorporation of N-14 at undesired sites. Taken together, this study represents the first application of selective unlabeling for sequence specific resonance assignments and opens up new avenues to using this methodology in protein structural studies

Hidden Giants in JWST's PEARLS: An Ultramassive z = 4.26 Submillimeter Galaxy that Is Invisible to HST

We present a multiwavelength analysis using the Submillimeter Array (SMA), James Clerk Maxwell Telescope, NOEMA, JWST, the Hubble Space Telescope (HST), and the Spitzer Space Telescope of two dusty strongly star-forming galaxies, 850.1 and 850.2, seen through the massive cluster lens A 1489. These SMA-located sources both lie at z = 4.26 and have bright dust continuum emission, but 850.2 is a UV-detected Lyman-break galaxy, while 850.1 is undetected at ≲ 2 μm, even with deep JWST/NIRCam observations. We investigate their stellar, interstellar medium, and dynamical properties, including a pixel-level spectral energy distribution analysis to derive subkiloparsec-resolution stellar-mass and A V maps. We find that 850.1 is one of the most massive and highly obscured, A V ∼ 5, galaxies known at z > 4 with M * ∼1011.8 M ⊙ (likely forming at z > 6), and 850.2 is one of the least massive and least obscured, A V ∼ 1, members of the z > 4 dusty star-forming population. The diversity of these two dust-mass-selected galaxies illustrates the incompleteness of galaxy surveys at z ≳ 3–4 based on imaging at ≲ 2 μm, the longest wavelengths feasible from HST or the ground. The resolved mass map of 850.1 shows a compact stellar-mass distribution, Remass ∼1 kpc, but its expected evolution means that it matches both the properties of massive, quiescent galaxies at z ∼ 1.5 and ultramassive early-type galaxies at z ∼ 0. We suggest that 850.1 is the central galaxy of a group in which 850.2 is a satellite that will likely merge in the near future. The stellar morphology of 850.1 shows arms and a linear bar feature that we link to the active dynamical environment it resides within

Magellanic system stars identified in SMACS J0723.3-7327 James Webb Space Telescope early release observations images

We identify 71 distant stars in James Webb Space Telescope/NIRCam early release observations (ERO) images of the field of galaxy cluster SMACS J0723.3-7327 (SMACS 0723). Given the relatively small (∼10°) angular separation between SMACS 0723 and the Large Magellanic Cloud (LMC), it is likely that these stars are associated with the LMC outskirts or the Leading Arm. This is further bolstered by a spectral energy distribution (SED) analysis, which suggests an excess of stars at a physical distance of 40–100 kpc, consistent with being associated with or located behind the Magellanic system. In particular, we find that the overall surface density of stars brighter than 27.0 mag in the field of SMACS 0723 is ∼2.3 times that of stars in a blank field with similar Galactic latitude (the North Ecliptic Pole Time Domain Field), and that the density of stars in the SMACS 0723 field with SED-derived distances consistent with the Magellanic system is ∼6.1 times larger than that of the blank field. The candidate stars at these distances are consistent with a stellar population at the same distance modulus with [Fe/H] = −1.0 and an age of ∼5.0 Gyr. On the assumption that all of the 71 stars are associated with the LMC, then the stellar density of the LMC at the location of the SMACS 0723 field is ∼740 stars kpc−3, which helps trace the density of stars in the LMC outskirts.J.S. acknowledges support from an undergraduate Arizona NASA Space Grant, Cooperative Agreement 80NSSC20M0041. R.A.W., S.H.C., and R.A.J. acknowledge support from NASA JWST Interdisciplinary Scientist grants NAG5-12460, NNX14AN10G and 80NSSC18K0200 from GSFC. C.J.C. and N.J.A. acknowledge support from the European Research Council (ERC) Advanced Investigator Grant EPOCHS (788113). M.A.M. acknowledges the support of a National Research Council of Canada Plaskett Fellowship, and the Australian Research Council Centre of Excellence for All Sky Astrophysics in 3 Dimensions (ASTRO 3D), through project number CE17010001. C.N.A.W. acknowledges funding from the JWST/NIRCam contract NASS-0215 to the University of Arizona. This work is based on observations made with the NASA/ESA/CSA James Webb Space Telescope. The data were obtained from the Mikulski Archive for Space Telescopes (MAST) at the Space Telescope Science Institute, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS 5-03127 for JWST. These observations are associated with JWST programs 1176, 2736, and 2738. All of the JWST data used in this paper can be found on doi:10.17909/b7hf-he53. This work is based on observations taken by the RELICS Treasury Program (Coe 2016, GO 14096) with the NASA/ESA HST, which is operated by the Association of Universities for Research in Astronomy, Inc., under NASA contract NAS5-26555. This work has made use of data from the European Space Agency (ESA) mission Gaia (https://www.cosmos.esa.int/gaia), processed by the Gaia Data Processing and Analysis Consortium (DPAC, https://www.cosmos.esa.int/web/gaia/dpac/consortium). Funding for the DPAC has been provided by national institutions, in particular the institutions participating in the Gaia Multilateral Agreement. We also acknowledge the JWST ERO team responsible for providing these products.Peer reviewe

PEARLS: A potentially isolated quiescent dwarf galaxy with a tip of the red giant branch distance of 30 Mpc

A wealth of observations have long suggested that the vast majority of isolated classical dwarf galaxies (M* = 107–109M⊙) are currently star forming. However, recent observations of the large abundance of "ultra-diffuse galaxies" beyond the reach of previous large spectroscopic surveys suggest that our understanding of the dwarf galaxy population may be incomplete. Here we report the serendipitous discovery of an isolated quiescent dwarf galaxy in the nearby Universe, which was imaged as part of the JWST PEARLS Guaranteed Time Observation program. Remarkably, individual red-giant branch stars are visible in this near-IR imaging, suggesting a distance of 30 ± 4 Mpc, and a wealth of archival photometry point to an sSFR of 2 × 10−11 yr−1 and star formation rate of 4 × 10−4M⊙ yr−1. Spectra obtained with the Lowell Discovery Telescope find a recessional velocity consistent with the Hubble Flow and >1500 km s−1 separated from the nearest massive galaxy in Sloan Digital Sky Survey suggesting that this galaxy was either quenched from internal mechanisms or had a very high-velocity (≳1000 km s−1) interaction with a nearby massive galaxy in the past. This analysis highlights the possibility that many nearby quiescent dwarf galaxies are waiting to be discovered and that JWST has the potential to resolve them.T.M.C. is grateful for support from the Beus Center for Cosmic Foundations. R.A.W., S.H.C., and R.A.J. acknowledge support from NASA JWST Interdisciplinary Scientist grants NAG5-12460, NNX14AN10G and 80NSSC18K0200 from GSFC. J.M.D. acknowledges the support of project PGC2018-101814-B-100 (MCIU/AEI/MINECO/FEDER, UE) Ministerio de Ciencia, Investigación y Universidades. This project was funded by the Agencia Estatal de Investigación, Unidad de Excelencia María de Maeztu, ref. MDM-2017-0765. C.C. is supported by the National Natural Science Foundation of China, Nos. 11803044, 11933003, 12173045. This work is sponsored (in part) by the Chinese Academy of Sciences (CAS), through a grant to the CAS South America Center for Astronomy (CASSACA). We acknowledge the science research grants from the China Manned Space Project with No. CMS-CSST-2021-A05. R.A.B. gratefully acknowledges support from the European Space Agency (ESA) Research Fellowship. C.J.C. acknowledges support from the European Research Council (ERC) Advanced Investigator Grant EPOCHS (788113). C.N.A.W. acknowledges funding from the JWST/NIRCam contract NASS-0215 to the University of Arizona. M.A.M. acknowledges the support of a National Research Council of Canada Plaskett Fellowship, and the Australian Research Council Centre of Excellence for All Sky Astrophysics in 3 Dimensions (ASTRO 3D), through project number CE17010001.Peer reviewe

MIL Cities and MIL Citizens: Informed, Engaged, Empowered by Media and Information Literacy (MIL).

Libro completoThe UNESCO UNITWIN Cooperation Programme on Media and Information Literacy and Intercultural Dialogue (MILID) is based on an initiative from the United Nations Educational, Scientific and Cultural Organization (UNESCO) and the UN Alliance of Civilizations (UNAOC). This Network was created in line with UNESCO’s mission and objectives, as well as the mandate of UNAOC, to serve as a catalyst and facilitator helping to give impetus to innovative projects aimed at reducing polarization among nations and cultures through mutual partnerships. This UNITWIN Network is composed of universities from different geographical areas: Autonomous University of Barcelona (Spain), University of the West Indies (Jamaica), Cairo University (Egypt), University of Sao Paulo (Brazil), Temple University (USA), Tsinghua University (China), Moulay Ismail University (Morocco), Sidi Mohamed Ben Abdellah University (Morocco), University of Guadalajara (Mexico), Western University (Canada), University of Gothenburg (Sweden), Sorbonne Nouvelle University (France), Punjabi University, Patiala (India), University of the South Pacific (Fiji), University of South Africa (South Africa), Nnamdi Azikiwe University (Nigeria), Ahmadu Bello University (Nigeria), Lagos State University (Nigeria), University of Jors (Nigeria), University of Calabar (Nigeria), Hosei University (Japan), University of Latvia (Latvia), Moscow Pedagogical State University (Russia), Corporación Universitaria Minuto de Dios UNIMINUTO (Colombia), Vytautas Magnus University (Lithuania), MICA (India), University of Campinas (Brazil). The main objectives of the Network are to foster collaboration among member universities, to build capacity in each of the countries in order to empower them to advance media and information literacy and intercultural dialogue, and to promote freedom of speech, freedom of information and the free flow of ideas and knowledge. Specific objectives include acting as an observatory for the role of media and information literacy (MIL) in promoting civic participation, democracy and development as well as enhancing intercultural and cooperative research on MIL. The programme also aims at promoting global actions related to MIL and intercultural dialogue. In such a context, a MILID Yearbook series is an important initiative. This MILID Yearbook is a result of a collaboration between UNESCO UNITWIN Cooperation Programme on Media and Information Literacy and Intercultural Dialogue, The Corporación Universitaria Minuto de Dios - UNIMINUTO (Colombia) and the University of Gothenburg (Sweden)

Unfolding of diphtheria toxin: identification of hydrophobic sites exposed on lowering of pH by photolabeling

We report here the use of a hydrophobic photoactivable reagent, 2-[3H]diazofluorene (DAF), to map the hydrophobic sites exposed when the pH is lowered in diphtheria toxin (DT). The reagent binds to DT, and on photolysis with light of wavelength >350 nm, it covalently attaches itself to DT. The labeling was observed to increase considerably when the pH was lowered from 7.4 to 5.2. Although both A- and B-chains were labeled to a similar degree at pH 7.4, at lower pH (5.2), B-chain was labeled to a much higher extent. Subsequent chemical and enzymatic fragmentation of DT followed by separation indicated that the putative transmembrane domain was labeled to its maximum extent at pH 5.2, with the bulk of labeling associated with residues 340-459. Protein sequencing analysis indicated that the two buried hydrophobic helices, identified in the crystal structure and suggested to insert and span the membrane bilayer, corresponding to residues 326–347 and 358–376, are strongly labeled. The Pro-345 residue was observed to be labeled maximally at lower pH values. Finally, the DAF labeling pattern indicated that the parent structural motifs are retained at low pH, suggesting that the low pH conformation of DT corresponds to an equilibrium molten globule state

Magellanic System Stars Identified in SMACS J0723.3-7327 James Webb Space Telescope Early Release Observations Images

We identify 71 distant stars in James Webb Space Telescope/NIRCam early release observations (ERO) images of the field of galaxy cluster SMACS J0723.3-7327 (SMACS 0723). Given the relatively small (∼10°) angular separation between SMACS 0723 and the Large Magellanic Cloud (LMC), it is likely that these stars are associated with the LMC outskirts or the Leading Arm. This is further bolstered by a spectral energy distribution (SED) analysis, which suggests an excess of stars at a physical distance of 40–100 kpc, consistent with being associated with or located behind the Magellanic system. In particular, we find that the overall surface density of stars brighter than 27.0 mag in the field of SMACS 0723 is ∼2.3 times that of stars in a blank field with similar Galactic latitude (the North Ecliptic Pole Time Domain Field), and that the density of stars in the SMACS 0723 field with SED-derived distances consistent with the Magellanic system is ∼6.1 times larger than that of the blank field. The candidate stars at these distances are consistent with a stellar population at the same distance modulus with [Fe/H] = −1.0 and an age of ∼5.0 Gyr. On the assumption that all of the 71 stars are associated with the LMC, then the stellar density of the LMC at the location of the SMACS 0723 field is ∼740 stars kpc ^−3 , which helps trace the density of stars in the LMC outskirts