Evidence for an Essential Deglycosylation-Independent Activity of PNGase in Drosophila melanogaster

BACKGROUND: Peptide:N-glycanase (PNGase) is an enzyme which releases N-linked glycans from glycopeptides/glycoproteins. This enzyme plays a role in the ER-associated degradation (ERAD) pathway in yeast and mice, but the biological importance of this activity remains unknown. PRINCIPAL FINDINGS: In this study, we characterized the ortholog of cytoplasmic PNGases, PNGase-like (Pngl), in Drosophila melanogaster. Pngl was found to have a molecular weight of approximately 74K and was mainly localized in the cytosol. Pngl lacks a CXXC motif that is critical for enzymatic activity in other species and accordingly did not appear to possess PNGase activity, though it still retains carbohydrate-binding activity. We generated microdeletions in the Pngl locus in order to investigate the functional importance of this protein in vivo. Elimination of Pngl led to a serious developmental delay or arrest during the larval and pupal stages, and surviving mutant adult males and females were frequently sterile. Most importantly, these phenotypes were rescued by ubiquitous expression of Pngl, clearly indicating that those phenotypic consequences were indeed due to the lack of functional Pngl. Interestingly, a putative "catalytic-inactive" mutant could not rescue the growth-delay phenotype, indicating that a biochemical activity of this protein is important for its biological function. CONCLUSION: Pngl was shown to be inevitable for the proper developmental transition and the biochemical properties other than deglycosylation activity is important for its biological function

Two-Loop Quark Self-Energy in a New Formalism (II): Renormalization of the Quark Propagator in the Light-Cone Gauge

The complete two-loop correction to the quark propagator, consisting of the spider, rainbow, gluon bubble and quark bubble diagrams, is evaluated in the noncovariant light-cone gauge (lcg). (The overlapping self-energy diagram had already been computed.) The chief technical tools include the powerful matrix integration technique, the n^*-prescription for the spurious poles of 1/qn, and the detailed analysis of the boundary singularities in five- and six-dimensional parameter space. It is shown that the total divergent contribution to the two-loop correction Sigma_2 contains both covariant and noncovariant components, and is a local function of the external momentum p, even off the mass-shell, as all nonlocal divergent terms cancel exactly. Consequently, both the quark mass and field renormalizations are local. The structure of Sigma_2 implies a quark mass counterterm of the form $\delta m (lcg) = m\tilde\alpha_s C_F(3+\tilde\alpha_sW) + {\rm O} (\tilde\alpha_s^3)$, \tilde\alpha_s = g^2\Gamma(\eps)(4\pi)^{\eps -2}, with W depending only on the dimensional regulator epsilon, and on the numbers of colors and flavors. It turns out that \delta m(lcg) is identical to the mass counterterm in the general linear covariant gauge. Our results are in agreement with the Bassetto-Dalbosco-Soldati renormalization scheme.Comment: 36 pages Latex, 5 eps figures, to appear in Nucl.Phys.

Kinetic oxygen measurements by CVC96 in L-929 cell cultures

Generally animal and human cells use oxygen during their whole life. Consequently the oxygen use is a simple indicator to test the vitality of cells. When the vitality decreases by the delivery of toxic substances the decrease can be observed directly by the oxygen-use of the cells. To get fast information of the vitality of cells we have measured the O(2)-tension by testing a new model of a bioreactor, the Cell Vitality Checker 96 (CVC96), in practical application. With this CVC96, soon a simple test will exist for the measurement of the oxygen use. In this respect the question had to be answered whether the use in the laboratory is easy and whether oxygen as a parameter in the vitality test can also be applied in future for problems in the field of material testing

Predicting phase equilibria in polydisperse systems

Many materials containing colloids or polymers are polydisperse: They comprise particles with properties (such as particle diameter, charge, or polymer chain length) that depend continuously on one or several parameters. This review focusses on the theoretical prediction of phase equilibria in polydisperse systems; the presence of an effectively infinite number of distinguishable particle species makes this a highly nontrivial task. I first describe qualitatively some of the novel features of polydisperse phase behaviour, and outline a theoretical framework within which they can be explored. Current techniques for predicting polydisperse phase equilibria are then reviewed. I also discuss applications to some simple model systems including homopolymers and random copolymers, spherical colloids and colloid-polymer mixtures, and liquid crystals formed from rod- and plate-like colloidal particles; the results surveyed give an idea of the rich phenomenology of polydisperse phase behaviour. Extensions to the study of polydispersity effects on interfacial behaviour and phase separation kinetics are outlined briefly.Comment: 48 pages, invited topical review for Journal of Physics: Condensed Matter; uses Institute of Physics style file iopart.cls (included

Atmospheric Heating and Wind Acceleration: Results for Cool Evolved Stars based on Proposed Processes

A chromosphere is a universal attribute of stars of spectral type later than ~F5. Evolved (K and M) giants and supergiants (including the zeta Aurigae binaries) show extended and highly turbulent chromospheres, which develop into slow massive winds. The associated continuous mass loss has a significant impact on stellar evolution, and thence on the chemical evolution of galaxies. Yet despite the fundamental importance of those winds in astrophysics, the question of their origin(s) remains unsolved. What sources heat a chromosphere? What is the role of the chromosphere in the formation of stellar winds? This chapter provides a review of the observational requirements and theoretical approaches for modeling chromospheric heating and the acceleration of winds in single cool, evolved stars and in eclipsing binary stars, including physical models that have recently been proposed. It describes the successes that have been achieved so far by invoking acoustic and MHD waves to provide a physical description of plasma heating and wind acceleration, and discusses the challenges that still remain.Comment: 46 pages, 9 figures, 1 table; modified and unedited manuscript; accepted version to appear in: Giants of Eclipse, eds. E. Griffin and T. Ake (Berlin: Springer

Ultrafast Structural Dynamics of BlsA, a Photoreceptor from the Pathogenic Bacterium Acinetobacter baumannii

Acinetobacter baumannii is an important human pathogen that can form biofilms and persist under harsh environmental conditions. Biofilm formation and virulence are modulated by blue light, which is thought to be regulated by a BLUF protein, BlsA. To understand the molecular mechanism of light sensing, we have used steady-state and ultrafast vibrational spectroscopy to compare the photoactivation mechanism of BlsA to the BLUF photosensor AppA from Rhodobacter sphaeroides. Although similar photocycles are observed, vibrational data together with homology modeling identify significant differences in the β5 strand in BlsA caused by photoactivation, which are proposed to be directly linked to downstream signaling