101 research outputs found
Crystallographic and Spectroscopic Snapshots Reveal a Dehydrogenase in Action
Aldehydes are ubiquitous intermediates in metabolic pathways and their innate reactivity can often make them quite unstable. There are several aldehydic intermediates in the metabolic pathway for tryptophan degradation that can decay into neuroactive compounds that have been associated with numerous neurological diseases. An enzyme of this pathway, 2-aminomuconate-6-semialdehyde dehydrogenase, is responsible for âdisarmingâ the final aldehydic intermediate. Here we show the crystal structures of a bacterial analogue enzyme in five catalytically relevant forms: resting state, one binary and two ternary complexes, and a covalent, thioacyl intermediate. We also report the crystal structures of a tetrahedral, thiohemiacetal intermediate, a thioacyl intermediate and an NADĂŸ-bound complex from an active site mutant. These covalent intermediates are characterized by single-crystal and solution-state electronic absorption spectroscopy. The crystal structures reveal that the substrate undergoes an E/Z isomerization at the enzyme active site before an sp3-to-sp2 transition during enzyme-mediated oxidation
The emergence of convergence
Science is increasingly a collaborative pursuit. Although the modern scientific enterprise owes much to individuals working at the core of their field, humanity is increasingly confronted by highly complex problems that require the integration of a variety of disciplinary and methodological expertise. In 2016, the U.S. National Science Foundation launched an initiative prioritizing support for convergence research as a means of âsolving vexing research problems, in particular, complex problems focusing on societal needs.â We discuss our understanding of the objectives of convergence research and describe in detail the conditions and processes likely to generate successful convergence research. We use our recent experience as participants in a convergence workshop series focused on resilience in the Arctic to highlight key points. The emergence of resilience science over the past 50 years is presented as a successful contemporary example of the emergence of convergence. We close by describing some of the challenges to the development of convergence research, such as timescales and discounting the future, appropriate metrics of success, allocation issues, and funding agency requirements
Acoustic transfer of protein crystals from agarose pedestals to micromeshes for high-throughput screening
An acoustic high-throughput screening method is described for harvesting protein crystals and combining the protein crystals with chemicals such as a fragment library
The Structure Of The Giant Haemoglobin From Glossoscolex Paulistus.
The sequences of all seven polypeptide chains from the giant haemoglobin of the free-living earthworm Glossoscolex paulistus (HbGp) are reported together with the three-dimensional structure of the 3.6 MDa complex which they form. The refinement of the full particle, which has been solved at 3.2 Ă
resolution, the highest resolution reported to date for a hexagonal bilayer haemoglobin composed of 12 protomers, is reported. This has allowed a more detailed description of the contacts between subunits which are essential for particle stability. Interpretation of features in the electron-density maps suggests the presence of metal-binding sites (probably Zn(2+) and Ca(2+)) and glycosylation sites, some of which have not been reported previously. The former appear to be important for the integrity of the particle. The crystal structure of the isolated d chain (d-HbGp) at 2.1 Ă
resolution shows different interchain contacts between d monomers compared with those observed in the full particle. Instead of forming trimers, as seen in the complex, the isolated d chains associate to form dimers across a crystallographic twofold axis. These observations eliminate the possibility that trimers form spontaneously in solution as intermediates during the formation of the dodecameric globin cap and contribute to understanding of the possible ways in which the particle self-assembles.711257-127
Hitting the target: fragment screening with acoustic in situ co-crystallization of proteins plus fragment libraries on pin-mounted data-collection micromeshes
A method is presented for screening fragment libraries using acoustic droplet ejection to co-crystallize proteins and chemicals directly on micromeshes with as little as 2.5â
nl of each component. This method was used to identify previously unreported fragments that bind to lysozyme, thermolysin, and trypsin
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Photoreversible interconversion of a phytochrome photosensory module in the crystalline state.
A major barrier to defining the structural intermediates that arise during the reversible photointerconversion of phytochromes between their biologically inactive and active states has been the lack of crystals that faithfully undergo this transition within the crystal lattice. Here, we describe a crystalline form of the cyclic GMP phosphodiesterases/adenylyl cyclase/FhlA (GAF) domain from the cyanobacteriochrome PixJ in Thermosynechococcus elongatus assembled with phycocyanobilin that permits reversible photoconversion between the blue light-absorbing Pb and green light-absorbing Pg states, as well as thermal reversion of Pg back to Pb. The X-ray crystallographic structure of Pb matches previous models, including autocatalytic conversion of phycocyanobilin to phycoviolobilin upon binding and its tandem thioether linkage to the GAF domain. Cryocrystallography at 150 K, which compared diffraction data from a single crystal as Pb or after irradiation with blue light, detected photoconversion product(s) based on Fobs - Fobs difference maps that were consistent with rotation of the bonds connecting pyrrole rings C and D. Further spectroscopic analyses showed that phycoviolobilin is susceptible to X-ray radiation damage, especially as Pg, during single-crystal X-ray diffraction analyses, which could complicate fine mapping of the various intermediate states. Fortunately, we found that PixJ crystals are amenable to serial femtosecond crystallography (SFX) analyses using X-ray free-electron lasers (XFELs). As proof of principle, we solved by room temperature SFX the GAF domain structure of Pb to 1.55-Ă
resolution, which was strongly congruent with synchrotron-based models. Analysis of these crystals by SFX should now enable structural characterization of the early events that drive phytochrome photoconversion
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Spring 1958
Dedication of Turf Clippings to Robert Williams (1) Picture - Stockbridge Turf majors (2) Title page and contents (3) Greetings from Dean Sieling - College of Agriculture (4) Outstanding Men in Turfgrass Honored (4) Word From the Editor of Turf Clippings (5) Message From the 1958 Winter School President (6) Summary of 1958 University of Massachusetts Turfgrass Conference - Al Radko (6-9) Importance of Superintendences Associations - Anthony B. Caranci (10) Picture - Turf Grass Interest Stretch from Coast to Coast (11) Picture - 1958 Winter School for Turf Managers (12) Golf Courses in California - James C. Scott (13) The Constant Battle - Joseph Troll (13) Recognition of the Golf Course Superintendent - Frederick Bove (14-15) 1958 Winter School Comments (16) Maintenance of Insurance Grounds - George J. Moore Jr. (17-18) Don\u27t Get Caught Short - Bruce Silven (19-20) We Have had It - They Now Have it - These Two Shall Have it - Orville O. Clapper (21) The Golf Course Superintendent - Prof. L. S. Dickinson (22-23) Picture - Winter School Alumni Meet at Conference (24) Cartoons (25) Cost of Lawn and Golf Course Construction - Geoffrey Cornish (26-27) Turf Club News (28-29) 1957 Horticulture Show Winner (30) Frosting on the Cake (30) Number One Graduate (31) 1957 Stockbridge Turf majors - Work (32) Turf management Club Associate Memberships (32
The adaptability of the ion binding site by the Ag(I)/Cu(I) periplasmic chaperone SilF
The periplasmic chaperone SilF has been identified as part of an Ag(I) detoxification system in Gram negative bacteria. Sil proteins also bind Cu(I), but with reported weaker affinity, therefore leading to the designation of a specific detoxification system for Ag(I). Using isothermal titration calorimetry we show that binding of both ions is not only tighter than previously thought, but of very similar affinities. We investigated the structural origins of ion binding using molecular dynamics and QM/MM simulations underpinned by structural and biophysical experiments. The results of this analysis showed that the binding site adapts to accommodate either ion, with key interactions with the solvent in the case of Cu(I). The implications of this are that Gram negative bacteria do not appear to have evolved a specific Ag(I) efflux system but take advantage of the existing Cu(I) detoxification system. Therefore, there are consequences for how we define a particular metal resistance mechanism and understand its evolution in the environment
The 2001 Superoutburst of WZ Sagittae
We report the results of a worldwide campaign to observe WZ Sagittae during
its 2001 superoutburst. After a 23-year slumber at V=15.5, the star rose within
2 days to a peak brightness of 8.2, and showed a main eruption lasting 25 days.
The return to quiescence was punctuated by 12 small eruptions, of ~1 mag
amplitude and 2 day recurrence time; these "echo outbursts" are of uncertain
origin, but somewhat resemble the normal outbursts of dwarf novae. After 52
days, the star began a slow decline to quiescence.
Periodic waves in the light curve closely followed the pattern seen in the
1978 superoutburst: a strong orbital signal dominated the first 12 days,
followed by a powerful /common superhump/ at 0.05721(5) d, 0.92(8)% longer than
P_orb. The latter endured for at least 90 days, although probably mutating into
a "late" superhump with a slightly longer mean period [0.05736(5) d]. The
superhump appeared to follow familiar rules for such phenomena in dwarf novae,
with components given by linear combinations of two basic frequencies: the
orbital frequency omega_o and an unseen low frequency Omega, believed to
represent the accretion disk's apsidal precession. Long time series reveal an
intricate fine structure, with ~20 incommensurate frequencies. Essentially all
components occurred at a frequency n(omega_o)-m(Omega), with m=1, ..., n. But
during its first week, the common superhump showed primary components at n
(omega_o)-Omega, for n=1, 2, 3, 4, 5, 6, 7, 8, 9 (i.e., m=1 consistently); a
month later, the dominant power shifted to components with m=n-1. This may
arise from a shift in the disk's spiral-arm pattern, likely to be the
underlying cause of superhumps.
The great majority of frequency components ... . (etc., abstract continues)Comment: PDF, 54 pages, 4 tables, 21 figures, 1 appendix; accepted, in press,
to appear July 2002, PASP; more info at http://cba.phys.columbia.edu
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