Context-dependent effects of asparagine glycosylation on Pin WW folding kinetics and thermodynamics

Asparagine glycosylation is one of the most common and important post-translational modifications of proteins in eukaryotic cells. N-glycosylation occurs when a triantennary glycan precursor is transferred en bloc to a nascent polypeptide (harboring the N-X-T/S sequon) as the peptide is cotranslationally translocated into the endoplasmic reticulum (ER). In addition to facilitating binding interactions with components of the ER proteostasis network, N-glycans can also have intrinsic effects on protein folding by directly altering the folding energy landscape. Previous work from our laboratories (Hanson et al. Proc. Natl. Acad. Sci. U.S.A. 2009, 109, 3131-3136; Shental-Bechor, D.; Levy, Y. Proc. Natl. Acad. Sci. U.S.A. 2008, 105, 8256-8261) suggested that the three sugar residues closest to the protein are sufficient for accelerating protein folding and stabilizing the resulting structure in vitro; even a monosaccharide can have a dramatic effect. The highly conserved nature of these three proximal sugars in N-glycans led us to speculate that introducing an N-glycosylation site into a protein that is not normally glycosylated would stabilize the protein and increase its folding rate in a manner that does not depend on the presence of specific stabilizing protein-saccharide interactions. Here, we test this hypothesis experimentally and computationally by incorporating an N-linked GlcNAc residue at various positions within the Pin WW domain, a small β-sheet-rich protein. The results show that an increased folding rate and enhanced thermodynamic stability are not general, context-independent consequences of N-glycosylation. Comparison between computational predictions and experimental observations suggests that generic glycan-based excluded volume effects are responsible for the destabilizing effect of glycosylation at highly structured positions. However, this reasoning does not adequately explain the observed destabilizing effect of glycosylation within flexible loops. Our data are consistent with the hypothesis that specific, evolved protein-glycan contacts must also play an important role in mediating the beneficial energetic effects on protein folding that glycosylation can confer.     

Breaking away: violation of distortion emission phase-frequency invariance at low frequencies

The phase versus frequency function of the distortion product otoacoustic emission (DPOAE) at 2f(1) - f(2) is approximately invariant at frequencies above 1.5 kHz in human subjects when recorded with a constant f(2)/f(1). However, a secular break from this invariance has been observed at lower frequencies where the phase-gradient becomes markedly steeper. Apical DPOAEs, such as 2f(1)?- f(2), are known to contain contributions from multiple sources. This experiment asked whether the phase behavior of the ear canal DPOAE at low frequencies is driven by the phase of the component from the distortion product (DP) region at 2f(1)?- f(2), which exhibits rapid phase accumulation. Placing a suppressor tone close in the frequency to 2f(1)?- f(2) reduced the contribution of this component to the ear canal DPOAE in normal-hearing adult human ears. When the contribution of this component was reduced, the phase behavior of the ear canal DPOAE was not altered, suggesting that the breaking from DPOAE phase invariance at low frequencies is an outcome of apical-basal differences in cochlear mechanics. The deviation from DPOAE phase invariance appears to be a manifestation of the breaking from approximate scaling symmetry in the human cochlear apex.     

Emergent cooperation amongst competing agents in minority games

We study a variation of the minority game. There are N agents. Each has to choose between one of two alternatives every day, and there is a reward to each member of the smaller group. The agents cannot communicate with each other, but try to guess the choice others will make, based only on the past history of the number of people choosing the two alternatives. We describe a simple probabilistic strategy using which the agents, acting independently, and trying to maximize their individual expected payoff, still achieve a very efficient overall utilization of resources, and the average deviation of the number of happy agents per day from the maximum possible can be made O(N^?), for any ?>0. We also show that a single agent does not expect to gain by not following the strategy.     

NHC-catalysed diastereoselective synthesis of multifunctionalised piperidines via cascade reaction of enals with azalactones

NHC-catalysed azalactone ring-opening and piperidine ring-closing cascade with α,β-unsaturated aldehydes (enals) in a one-pot operation is reported. The present reaction cascade offers a convenient method for a highly diastereoselective synthesis of multifunctionalised piperidines in excellent yields under mild conditions.     

Infrared absorption and Raman spectroscopy studies of InSbBi layers grown by liquid phase epitaxy

We report on the results of optical absorption and Raman spectroscopy measurements on InSbBi layers grown by liquid phase technique. A maximum Bi content of 0.4 at.%, as measured by energy dispersive X-ray (EDX) technique, is used in the experiments. Optical absorption measurements made on the samples indicate a room temperature energy band gap reduction up to about 6 meV with respect to undoped InSb layers grown by the same technique. Bi content calculated from this band gap reduction agrees with that obtained from EDX. A weak peak obtained at 152 cm^?1 in the Raman spectrum of the material is identified with the longitudinal optical phonon mode of InBi. Further a mode at 140 cm^?1 is observed due to isolated Bi atoms at the interstitial sites.     

Fracture analysis of wheel hub fabricated from pressure die cast aluminum alloy

A catastrophic failure of wheel hub occurred during service. The nature of failure was a corner crack emanating from a spoke hole. An analytical investigation was carried out using the tool of Linear Elastic Fracture Mechanics to establish the cause of failure. The nonlinear behaviour is due to the presence of material inhomogeneities and micro-discontinuities. Plastic zone correction is made. The presence of material inhomogeneities and micro-discontinuities tend to alter the conditions near the crack tip. An analytical estimation was carried out in order to calculate the minimum number of cycles covered by the wheel hub in service.The initiation of crack growth is complex because of heterogeneity and morphology of the fracture surface. Fractographic and metallographic studies are carried out to assist the understanding of the corner cracking problem.     

A comparative study of distortion-product-otoacoustic-emission fine structure in human newborns and adults with normal hearing

Distortion product otoacoustic emissions (DPOAE) measured in human newborns are not adult-like. More than a decade of work from various investigators has created a well-developed body of evidence describing these differences but the putative anatomy or physiology has only been partially explained. Recently, Abdala and Keefe [J. Acoust. Soc. Am. 120, 3832-3842 (2006)] have identified outer and middle ear immaturities that at least partially describe the differences observed between newborn and adult input-output functions and suppression tuning curves. DPOAE fine structure characteristics and their maturation have not been examined to any extent in the literature. Fine structure characteristics in two groups of ten newborns and young adults with normal hearing sensitivity are compared here. Consistent with previous reports, the newborns show higher DPOAE levels; greater fine structure depth and wider fine structure spacing is also observed in the newborns. Differences in fine structure morphology are also observed between the two age groups. While some of these findings are attributable to an immature outer and middle ear system in the newborns, it is argued that some observed differences in fine structure characteristics might be due to remnant immaturities in passive motion of the basilar membrane in the newborn cochlea.     

Wettability, FTIR and dielectric studies of 1,4-dioxane and water system

Wettability studies are of importance for electronic devices. Various methods are known to convert the hydrophobic substrates to hydrophilic substrates, but the studies on the relative dependence of wettability with varying concentrations of an aqueous system are meager. The wetting of different substrates with varying concentration of 1,4-dioxane in water is investigated and the results of concentration dependence of wetting are presented. The FTIR spectrum shows a blue shift of the OH peak--a feature typical of aqueous-1,4-dioxane systems. Concentration dependence of dielectric permittivity of this system also showed an anomaly.