Evolving proteins at Darwin's bicentenary

A report of the Biochemical Society/Wellcome Trust meeting 'Protein Evolution - Sequences, Structures and Systems', Hinxton, UK, 26-27 January 2009

Derivative processes for modelling metabolic fluxes.

MOTIVATION: One of the challenging questions in modelling biological systems is to characterize the functional forms of the processes that control and orchestrate molecular and cellular phenotypes. Recently proposed methods for the analysis of metabolic pathways, for example, dynamic flux estimation, can only provide estimates of the underlying fluxes at discrete time points but fail to capture the complete temporal behaviour. To describe the dynamic variation of the fluxes, we additionally require the assumption of specific functional forms that can capture the temporal behaviour. However, it also remains unclear how to address the noise which might be present in experimentally measured metabolite concentrations. RESULTS: Here we propose a novel approach to modelling metabolic fluxes: derivative processes that are based on multiple-output Gaussian processes (MGPs), which are a flexible non-parametric Bayesian modelling technique. The main advantages that follow from MGPs approach include the natural non-parametric representation of the fluxes and ability to impute the missing data in between the measurements. Our derivative process approach allows us to model changes in metabolite derivative concentrations and to characterize the temporal behaviour of metabolic fluxes from time course data. Because the derivative of a Gaussian process is itself a Gaussian process, we can readily link metabolite concentrations to metabolic fluxes and vice versa. Here we discuss how this can be implemented in an MGP framework and illustrate its application to simple models, including nitrogen metabolism in Escherichia coli. AVAILABILITY AND IMPLEMENTATION: R code is available from the authors upon request

Derivative processes for modelling metabolic fluxes

Motivation: One of the challenging questions in modelling biological systems is to characterize the functional forms of the processes that control and orchestrate molecular and cellular phenotypes. Recently proposed methods for the analysis of metabolic pathways, for example, dynamic flux estimation, can only provide estimates of the underlying fluxes at discrete time points but fail to capture the complete temporal behaviour. To describe the dynamic variation of the fluxes, we additionally require the assumption of specific functional forms that can capture the temporal behaviour. However, it also remains unclear how to address the noise which might be present in experimentally measured metabolite concentrations. Results: Here we propose a novel approach to modelling metabolic fluxes: derivative processes that are based on multiple-output Gaussian processes (MGPs), which are a flexible non-parametric Bayesian modelling technique. The main advantages that follow from MGPs approach include the natural non-parametric representation of the fluxes and ability to impute the missing data in between the measurements. Our derivative process approach allows us to model changes in metabolite derivative concentrations and to characterize the temporal behaviour of metabolic fluxes from time course data. Because the derivative of a Gaussian process is itself a Gaussian process, we can readily link metabolite concentrations to metabolic fluxes and vice versa. Here we discuss how this can be implemented in an MGP framework and illustrate its application to simple models, including nitrogen metabolism in Escherichia coli

Some protein interaction data do not exhibit power law statistics

It has been claimed that protein-protein interaction (PPI) networks are scale-free based on the observation that the node degree sequence follows a power law. Here we argue that these claims are likely to be based on erroneous statistical analysis. Typically, the supporting data are presented using frequency-degree plots. We show that such plots can be misleading, and should correctly be replaced by rank-degree plots. We provide two PPI network examples in which the frequency-degree plots appear linear on a log-log scale, but the rank-degree plots demonstrate that the node degree sequence is far from a power law. We conclude that at least these PPI networks are not scale-free.Comment: 4 pages, 2 figure

Airborne fine-resolution UHF radar: an approach to the study of englacial reflections, firn compaction and ice attenuation rates

This is the published version. Copyright 2015 International Glaciological SocietyWe have built and operated an ultra-wideband UHF pulsed-chirp radar for measuring firn stratigraphy from airborne platforms over the ice sheets of Greenland and West Antarctica. Our analysis found a wide range of capabilities, including imaging of post firn–ice transition horizons and sounding of shallow glaciers and ice shelves. Imaging of horizons to depths exceeding 600 m was possible in the colder interior regions of the ice sheet, where scattering from the ice surface and inclusions was minimal. The radar's high sensitivity and large dynamic range point to loss tangent variations as the dominant mechanism for these englacial reflective horizons. The radar is capable of mapping interfaces with reflection coefficients as low as –80 dB near the firn–ice transition and as low as –64 dB at depths of 600 m. We found that firn horizon reflectivity strongly mirrored density variance, a result of the near-unity interfacial transmission coefficients. Zones with differing compaction mechanisms were also apparent in the data. We were able to sound many ice shelves and areas of shallow ice. We estimated ice attenuation rates for a few locations, and our attenuation estimates for the Ross Ice Shelf, West Antarctica, appear to agree well with earlier reported results

Identification of a Wide, Low-Mass Multiple System Containing the Brown Dwarf 2MASS J0850359+105716

We report our discovery of NLTT 20346 as an M5+M6 companion system to the tight binary (or triple) L dwarf 2MASS J0850359+105716. This nearby (~31 pc), widely separated (~7700 AU) quadruple system was identified through a cross-match of proper motion catalogs. Follow-up imaging and spectroscopy of NLTT 20346 revealed it to be a magnetically active M5+M6 binary with components separated by ~2" (50-80 AU). Optical spectroscopy of the components show only moderate Halpha emission corresponding to a statistical age of ~5 - 7 Gyr for both M dwarfs. However NLTT 20346 is associated with the XMM-Newton source J085018.9+105644, and based on X-ray activity the age of NLTT 20346 is between 250-450 Myr. Strong Li absorption in the optical spectrum of 2MASS J0850+1057 indicates an upper age limit of 0.8 - 1.5 Gyr favoring the younger age for the primary. Using evolutionary models in combination with an adopted system age of 0.25-1.5 Gyr indicates a total mass for 2MASS J0850+1057 of 0.07+/-0.02 Msun if it is a binary. NLTT 20346/2MASS J0850+1057 joins a growing list of hierarchical systems containing brown dwarf binaries and is among the lowest binding energy associations found in the field. Formation simulations via gravitational fragmentation of massive extended disks have successfully produced a specific analog to this system.Comment: 13 pages, accepted for publication to A

Discovery of unusual dimeric piperazyl cyclopeptides encoded by a Lentzea flaviverrucosa DSM 44664 biosynthetic supercluster

Rare actinomycetes represent an underexploited source of new bioactive compounds. Here, we report the use of a targeted metabologenomic approach to identify piperazyl compounds in the rare actinomycete Lentzea flaviverrucosa DSM 44664. These efforts to identify molecules that incorporate piperazate building blocks resulted in the discovery and structural elucidation of two dimeric biaryl-cyclohexapeptides, petrichorins A and B. Petrichorin B is a symmetric homodimer similar to the known compound chloptosin, but petrichorin A is unique among known piperazyl cyclopeptides because it is an asymmetric heterodimer. Due to the structural complexity of petrichorin A, solving its structure required a combination of several standard chemical methods plus in silico modeling, strain mutagenesis, and solving the structure of its biosynthetic intermediate petrichorin C for confident assignment. Furthermore, we found that the piperazyl cyclopeptides comprising each half of the petrichorin A heterodimer are made via two distinct nonribosomal peptide synthetase (NRPS) assembly lines, and the responsible NRPS enzymes are encoded within a contiguous biosynthetic supercluster on the L. flaviverrucosa chromosome. Requiring promiscuous cytochrome p450 crosslinking events for asymmetric and symmetric biaryl production, petrichorins A and B exhibited potent in vitro activity against A2780 human ovarian cancer, HT1080 fibrosarcoma, PC3 human prostate cancer, and Jurkat human T lymphocyte cell lines with IC50 values at low nM levels. Cyclic piperazyl peptides and their crosslinked derivatives are interesting drug leads, and our findings highlight the potential for heterodimeric bicyclic peptides such as petrichorin A for inclusion in future pharmaceutical design and discovery programs.Fil: Li, Chunshun. University Of Hawaii; Estados UnidosFil: Hu, Yifei. University Of Hawaii; Estados Unidos. Washington University in St. Louis; Estados UnidosFil: Wu, Xiaohua. University Of Hawaii; Estados UnidosFil: Stumpf, Spencer D.. Washington University in St. Louis; Estados UnidosFil: Qi, Yunci. Washington University in St. Louis; Estados UnidosFil: D'Alessandro, John M.. Washington University in St. Louis; Estados UnidosFil: Nepal, Keshav K.. Washington University in St. Louis; Estados UnidosFil: Sarotti, Ariel Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; ArgentinaFil: Cao, Shugeng. University Of Hawaii; Estados UnidosFil: Blodgett, Joshua A.V.. Washington University in St. Louis; Estados Unido

A large fraction of HLA class I ligands are proteasome-generated spliced peptides

The proteasome generates the epitopes presented on human leukocyte antigen (HLA) class I molecules that elicit CD8(+) T cell responses. Reports of proteasome-generated spliced epitopes exist, but they have been regarded as rare events. Here, however, we show that the proteasome-generated spliced peptide pool accounts for one-third of the entire HLA class I immunopeptidome in terms of diversity and one-fourth in terms of abundance. This pool also represents a unique set of antigens, possessing particular and distinguishing features. We validated this observation using a range of complementary experimental and bioinformatics approaches, as well as multiple cell types. The widespread appearance and abundance of proteasome-catalyzed peptide splicing events has implications for immunobiology and autoimmunity theories and may provide a previously untapped source of epitopes for use in vaccines and cancer immunotherapy

MemPrep, a new technology for isolating organellar membranes provides fingerprints of lipid bilayer stress

Biological membranes have a stunning ability to adapt their composition in response to physiological stress and metabolic challenges. Little is known how such perturbations affect individual organelles in eukaryotic cells. Pioneering work has provided insights into the subcellular distribution of lipids in the yeast Saccharomyces cerevisiae, but the composition of the endoplasmic reticulum (ER) membrane, which also crucially regulates lipid metabolism and the unfolded protein response, remains insufficiently characterized. Here, we describe a method for purifying organelle membranes from yeast, MemPrep. We demonstrate the purity of our ER membrane preparations by proteomics, and document the general utility of MemPrep by isolating vacuolar membranes. Quantitative lipidomics establishes the lipid composition of the ER and the vacuolar membrane. Our findings provide a baseline for studying membrane protein biogenesis and have important implications for understanding the role of lipids in regulating the unfolded protein response (UPR). The combined preparative and analytical MemPrep approach uncovers dynamic remodeling of ER membranes in stressed cells and establishes distinct molecular fingerprints of lipid bilayer stress.This work was funded by the VW foundation (Life?, #93089, #93092, #93090) to RE, MS, and JS, by the Deutsche Forschungsgemeinschaft in the framework of the SFB894 to RE and the SFB1027 to both JH and RE, and by the European Research Council under the European Union’s Horizon 2020 research and innovation program (grant agreement no. 866011) to RE. MS is an incumbent of the Dr. Gilbert Omenn and Martha Darling Professorial Chair in Molecular Genetics