Many peptide-derived natural products are produced by non-ribosomal peptide synthetases (NRPSs) in an assembly-line fashion. Each amino acid is coupled to a designated peptidyl carrier protein (PCP) through two distinct reactions catalysed sequentially by the single active site of the adenylation domain (A-domain). Accumulating evidence suggests that large-amplitude structural changes occur in different NRPS states; yet how these molecular machines orchestrate such biochemical sequences has remained elusive. Here, using single-molecule Forster resonance energy transfer, we show that the A-domain of gramicidin S synthetase I adopts structurally extended and functionally obligatory conformations for alternating between adenylation and thioester-formation structures during enzymatic cycles. Complementary biochemical, computational and small-angle X-ray scattering studies reveal interconversion among these three conformations as intrinsic and hierarchical where intra-A-domain organizations propagate to remodel inter-A-PCP didomain configurations during catalysis. The tight kinetic coupling between structural transitions and enzymatic transformations is quantified, and how the gramicidin S synthetase I A-domain utilizes its inherent conformational dynamics to drive directional biosynthesis with a flexibly linked PCP domain is revealed. Many natural products are produced by non-ribosomal peptide synthetases in an assembly-line fashion. How these molecular machines orchestrate the biochemical sequences has remained elusive. It is now understood that an extended-conformation ensemble is needed to coordinate chemical-transformation steps whereas the biosynthesis directionality is driven by the enzyme's innate conformational free energies.

We present a database resulting from high throughput experimentation, primarily on metal oxide solid state materials. The central relational database, the Materials Provenance Store (MPS), manages the metadata and experimental provenance from acquisition of raw materials, through synthesis, to a broad range of materials characterization techniques. Given the primary research goal of materials discovery of solar fuels materials, many of the characterization experiments involve electrochemistry, along with optical, structural, and compositional characterizations. The MPS is populated with all information required for executing common data queries, which typically do not involve direct query of raw data. The result is a database file that can be distributed to users so that they can independently execute queries and subsequently download the data of interest. We propose this strategy as an approach to manage the highly heterogeneous and distributed data that arises from materials science experiments, as demonstrated by the management of over 30 million experiments run on over 12 million samples in the present MPS release.

Scientific software registries and repositories improve software findability and research transparency, provide information for software citations, and foster preservation of computational methods in a wide range of disciplines. Registries and repositories play a critical role by supporting research reproducibility and replicability, but developing them takes effort and few guidelines are available to help prospective creators of these resources. To address this need, the FORCE11 Software Citation Implementation Working Group convened a Task Force to distill the experiences of the managers of existing resources in setting expectations for all stakeholders. In this article, we describe the resultant best practices which include defining the scope, policies, and rules that govern individual registries and repositories, along with the background, examples, and collaborative work that went into their development. We believe that establishing specific policies such as those presented here will help other scientific software registries and repositories better serve their users and their disciplines.

Scientific software registries and repositories improve software findability and research transparency, provide information for software citations, and foster preservation of computational methods in a wide range of disciplines.Registries and repositories play a critical role by supporting research reproducibility and replicability, but developing them takes effort and few guidelines are available to help prospective creators of these resources.To address this need, the FORCE11 Software Citation Implementation Working Group convened a Task Force to distill the experiences of the managers of existing resources in setting expectations for all stakeholders.In this paper, we describe the resultant best practices which include defining the scope, policies, and rules that govern individual registries and repositories, along with the background, examples, and collaborative work that went into their development.We believe that establishing specific policies such as those presented here will help other scientific software registries and repositories better serve their users and their disciplines.

Protein tyrosine phosphatase B (PtpB) from Mycobacterium tuberculosis (Mtb) extends the bacteria's survival in hosts and hence is a potential target for Mtb-specific drugs. To study how Mtb-specific sequence insertions in PtpB may regulate access to its active site through large-amplitude conformational changes, we performed free-energy calculations using an all-atom explicit solvent model. Corroborated by biochemical assays, the results show that PtpB's active site is controlled via an "either/or" compound conformational gating mechanism, an unexpected discovery that Mtb has evolved to bestow a single enzyme with such intricate logical operations. In addition to providing unprecedented insights for its active-site surroundings, the findings also suggest new ways of inactivating PtpB.

Software Carpentry is a non-profit volunteer organization that was established in 1998 to teach science researchers basic, essential computing skills using a compressed, hands-on workshop approach, and focusing on open-source platforms wherever possible. Usually lasting 2 or 3 days, Software Carpentry workshops focus on teaching the basics of the unix command line, programming (using R or Python), database operation (using SQLite) and version control (using Git or Mercurial). The movement has expanded to include Data Carpentry lessons and workshops focused on cleaning and analyzing research data (including using OpenRefine), Library Carpentry workshops targeted at programming tools librarians could use in their work, and brand-new Author Carpentry lessons targeted at tools that support modern writing and publishing, such at managing bibliog. data using BibTeX and other open-source solns., managing scholarly identification using ORCiD, and other topics. Three Caltech Librarians have completed the Instructor Training to become certified Instructors, and to date we have held two Carpentry workshops for the Caltech community. Discussion will include the philosophy behind the Carpentry instruction approach and a debrief of the workshops held, its relevance to chem. researchers and information professionals, and why this style of instruction has found a home at the Caltech Library.

Nonribosomal peptide synthetases (NRPSs) are multidomain enzyme templates for the synthesis of bioactive peptides. Large-scale conformational changes during peptide assembly are obvious from crystal structures, yet their dynamics and coupling to catalysis are poorly understood. We have designed an NRPS FRET sensor to monitor, in solution and in real time, the adoption of the productive transfer conformation between phenylalanine-binding adenylation (A) and peptidyl-carrier-protein domains of gramicidin synthetase I from Aneurinibacillus migulanus. The presence of ligands, substrates or intermediates induced a distinct fluorescence resonance energy transfer (FRET) readout, which was pinpointed to the population of specific conformations or, in two cases, mixtures of conformations. A pyrophosphate switch and lysine charge sensors control the domain alternation of the A domain. The phenylalanine-thioester and phenylalanine-AMP products constitute a mechanism of product inhibition and release that is involved in ordered assembly-line peptide biosynthesis. Our results represent insights from solution measurements into the conformational dynamics of the catalytic cycle of NRPSs.

Protein conformational changes are known to play important roles in assorted biochemical and biological processes. Driven by thermal motions of surrounding solvent molecules, such a structural remodeling often occurs stochastically. Yet, regardless of how random the conformational reconfiguration may appear, it could in principle be described by a linear combination of a set of orthogonal modes which, in turn, are contained in the intramolecular distance distributions. The central challenge is how to obtain the distribution. This contribution proposes a Bayesian data-augmentation scheme to extract the predominant modes from only few distance distributions, be they from computational sampling or directly from experiments such as single-molecule Förster-type resonance energy transfer (smFRET). The inference of the complete protein structure from insufficient data was recognized as isomorphic to the missing-data problem in Bayesian statistical learning. Using smFRET data as an example, the missing coordinates were deduced, given protein structural constraints and multiple but limited number of smFRET distances; the Boltzmann weighing of each inferred protein structure was then evaluated using computational modeling to numerically construct the posterior density for the global protein conformation. The conformational modes were then determined from the iteratively converged overall conformational distribution using principal component analysis. Two examples were presented to illustrate these basic ideas as well as their practical implementation. The scheme described herein was based on the theory behind the powerful Tanner-Wang algorithm that guarantees convergence to the true posterior density. However, instead of assuming a mathematical model to calculate the likelihood as in conventional statistical inference, here the protein structure was treated as a statistical parameter and was imputed from the numerical likelihood function based on structural information, a probability model-free method. The framework put forth here is anticipated to be generally applicable, offering a new way to articulate protein conformational changes in a quantifiable manner.

Mycobacterium tuberculosis protein tyrosine phosphatase B (PtpB), which plays a key role in tuberculosis virulence, protects its active site from oxidation through a large-amplitude conformational change. The proposed regulatory mechanism for this protective motion involves a local unfolding event. We quantitatively show that local unfolding transitions can be coupled to other conformational changes in proteins. Molecular dynamics simulations were used to determine the conformations of a disordered protein region and the energy landscape for local unfolding. The accuracy of this energy landscape was tested and confirmed experimentally, providing rigorous support for our simulation approaches. Our work demonstrates that local unfolding can depend on protein conformation by changes in the stability of the disordered region. These results provide a mechanism for the internal regulation of protein conformational changes, which highlights new targets for the development of tuberculosis therapeutics.

Quantum dots are an excellent resource for demonstrating quantum phenomena. Two new methods for synthesizing quantum dots are presented. Proceeding at relatively low reaction temperatures, these exercises are safe and easy to conduct in an undergraduate student laboratory. The quantum dots prepared from the first method exhibited visible luminescence across a broad range of colors. The size-dependent spectral properties of quantum dots were examined quantitatively in the second method. Following this procedure, students in an advanced chemistry laboratory course synthesized their own quantum dots and gained experience with these important nanomaterials.

An efficient mixed molecular dynamics/quantum mechanics model has been applied to the water cluster system. The use of the MP2 method and correlation consistent basis sets, with appropriate correction for BSSE, allows for the accurate calculation of electronic and free energies for the formation of clusters of 2-10 water molecules. This approach reveals new low energy conformers for (H(2)O)(n=7,9,10). The water heptamer conformers comprise five different structural motifs ranging from a three-dimensional prism to a quasi-planar book structure. A prism-like structure is favored energetically at low temperatures, but a chair-like structure is the global Gibbs free energy minimum past 200 K. The water nonamers exhibit less complexity with all the low energy structures shaped like a prism. The decamer has 30 conformers that are within 2 kcal/mol of the Gibbs free energy minimum structure at 298 K. These structures are categorized into four conformer classes, and a pentagonal prism is the most stable structure from 0 to 320 K. Results can be used as benchmark values for empirical water models and density functionals, and the method can be applied to larger water clusters.

A mixed molecular dynamics/quantum mechanics model has been applied to the ammonium/water clustering system. The use of the high level MP2 calculation method and correlated basis sets, such as aug-cc-pVDZ and aug-cc-pVTZ, lends confidence in the accuracy of the extrapolated energies. These calculations provide electronic and free energies for the formation of clusters of ammonium and 1-10 water molecules at two different temperatures. Structures and thermodynamic values are in good agreement with previous experimental and theoretical results. The estimated concentration of these clusters in the troposphere was calculated using atmospheric amounts of ammonium and water. Results show the favorability of forming these clusters and implications for ion-induced nucleation in the atmosphere.