Lunzer Curated Biochemistry

Model-fit Km and performance (P = kcat/Km) against NAD and NADP, together with preference (PNAD/PNADP) for each allele

Induced Fit and the Catalytic Mechanism of Isocitrate Dehydrogenase

NADP⁺ dependent isocitrate dehydrogenase (IDH; EC 1.1.1.42) belongs to a large family of α-hydroxyacid oxidative β-decarboxylases that catalyze similar three-step reactions, with dehydrogenation to an oxaloacid intermediate preceding β-decarboxylation to an enol intermediate followed by tautomerization to the final α-ketone product. A comprehensive view of the induced fit needed for catalysis is revealed on comparing the first “fully closed” crystal structures of a <i>pseudo-</i>Michaelis complex of wild-type <i>Escherichia coli</i> IDH (<i>Eco</i>IDH) and the “fully closed” reaction product complex of the K100M mutant with previously obtained “quasi-closed” and “open” conformations. Conserved catalytic residues, binding the nicotinamide ring of NADP⁺ and the metal-bound substrate, move as rigid bodies during domain closure by a hinge motion that spans the central β-sheet in each monomer. Interactions established between Thr105 and Ser113, which flank the “phosphorylation loop”, and the nicotinamide mononucleotide moiety of NADP⁺ establish productive coenzyme binding. Electrostatic interactions of a Lys100-Leu103-Asn115-Glu336 tetrad play a pivotal role in assembling a catalytically competent active site. As predicted, Lys230* is positioned to deprotonate/reprotonate the α-hydroxyl in both reaction steps and Tyr160 moves into position to protonate C3 following β-decarboxylation. A proton relay from the catalytic triad Tyr160-Asp307-Lys230* connects the α-hydroxyl of isocitrate to the bulk solvent to complete the picture of the catalytic mechanism

DataSheet_1_Rolling Deck to Repository: Supporting the marine science community with data management services from academic research expeditions.docx

Direct observations of the oceans acquired on oceanographic research ships operated across the international community support fundamental research into the many disciplines of ocean science and provide essential information for monitoring the health of the oceans. A comprehensive knowledge base is needed to support the responsible stewardship of the oceans with easy access to all data acquired globally. In the United States, the multidisciplinary shipboard sensor data routinely acquired each year on the fleet of coastal, regional and global ranging vessels supporting academic marine research are managed by the Rolling Deck to Repository (R2R, rvdata.us) program. With over a decade of operations, the R2R program has developed a robust routinized system to transform diverse data contributions from different marine data providers into a standardized and comprehensive collection of global-ranging observations of marine atmosphere, ocean, seafloor and subseafloor properties that is openly available to the international research community. In this article we describe the elements and framework of the R2R program and the services provided. To manage all expeditions conducted annually, a fleet-wide approach has been developed using data distributions submitted from marine operators with a data management workflow designed to maximize automation of data curation. Other design goals are to improve the completeness and consistency of the data and metadata archived, to support data citability, provenance tracking and interoperable data access aligned with FAIR (findable, accessible, interoperable, reusable) recommendations, and to facilitate delivery of data from the fleet for global data syntheses. Findings from a collection-level review of changes in data acquisition practices and quality over the past decade are presented. Lessons learned from R2R operations are also discussed including the benefits of designing data curation around the routine practices of data providers, approaches for ensuring preservation of a more complete data collection with a high level of FAIRness, and the opportunities for homogenization of datasets from the fleet so that they can support the broadest re-use of data across a diverse user community.</p

Whole-Genome Analyses of Korean Native and Holstein Cattle Breeds by Massively Parallel Sequencing

<div><p>A main goal of cattle genomics is to identify DNA differences that account for variations in economically important traits. In this study, we performed whole-genome analyses of three important cattle breeds in Korea—Hanwoo, Jeju Heugu, and Korean Holstein—using the Illumina HiSeq 2000 sequencing platform. We achieved 25.5-, 29.6-, and 29.5-fold coverage of the Hanwoo, Jeju Heugu, and Korean Holstein genomes, respectively, and identified a total of 10.4 million single nucleotide polymorphisms (SNPs), of which 54.12% were found to be novel. We also detected 1,063,267 insertions–deletions (InDels) across the genomes (78.92% novel). Annotations of the datasets identified a total of 31,503 nonsynonymous SNPs and 859 frameshift InDels that could affect phenotypic variations in traits of interest. Furthermore, genome-wide copy number variation regions (CNVRs) were detected by comparing the Hanwoo, Jeju Heugu, and previously published Chikso genomes against that of Korean Holstein. A total of 992, 284, and 1881 CNVRs, respectively, were detected throughout the genome. Moreover, 53, 65, 45, and 82 putative regions of homozygosity (ROH) were identified in Hanwoo, Jeju Heugu, Chikso, and Korean Holstein respectively. The results of this study provide a valuable foundation for further investigations to dissect the molecular mechanisms underlying variation in economically important traits in cattle and to develop genetic markers for use in cattle breeding.</p></div