Structure and in silico simulations of a cold-active esterase reveals its prime cold-adaptation mechanism

Here we determined the structure of a cold active family IV esterase (EstN7) cloned from Bacillus cohnii strain N1. EstN7 is a dimer with a classical α/β hydrolase fold. It has an acidic surface that is thought to play a role in cold-adaption by retaining solvation under changed water solvent entropy at lower temperatures. The conformation of the functionally important cap region is significantly different to EstN7's closest relatives, forming a bridge-like structure with reduced helical content providing greater access to the active site through more than one substrate access tunnel. However, dynamics do not appear to play a major role in cold adaption. Molecular dynamics at different temperatures, rigidity analysis, normal mode analysis and geometric simulations of motion confirm the flexibility of the cap region but suggest that the rest of the protein is largely rigid. Rigidity analysis indicates the distribution of hydrophobic tethers is appropriate to colder conditions, where the hydrophobic effect is weaker than in mesophilic conditions due to reduced water entropy. Thus, it is likely that increased substrate accessibility and tolerance to changes in water entropy are important for of EstN7's cold adaptation rather than changes in dynamics.

Data-Independent Acquisition-Based Proteome and Phosphoproteome Profiling Reveals Early Protein Phosphorylation and Dephosphorylation Events in Arabidopsis Seedlings upon Cold Exposure

Protein phosphorylation plays an important role in mediating signal transduction in cold response in plants. To better understand how plants sense and respond to the early temperature drop, we performed data-independent acquisition (DIA) method-based mass spectrometry analysis to profile the proteome and phosphoproteome of Arabidopsis seedlings upon cold stress in a time-course manner (10, 30 and 120 min of cold treatments). Our results showed the rapid and extensive changes at the phosphopeptide levels, but not at the protein abundance levels, indicating cold-mediated protein phosphorylation and dephosphorylation events. Alteration of over 1200 proteins at phosphopeptide levels were observed within 2 h of cold treatment, including over 140 kinases, over 40 transcriptional factors and over 40 E3 ligases, revealing the complexity of regulation of cold adaption. We summarized cold responsive phosphoproteins involved in phospholipid signaling, cytoskeleton reorganization, calcium signaling, and MAPK cascades. Cold-altered levels of 73 phosphopeptides (mostly novel cold-responsive) representing 62 proteins were validated by parallel reaction monitoring (PRM). In summary, this study furthers our understanding of the molecular mechanisms of cold adaption in plants and strongly supports that DIA coupled with PRM are valuable tools in uncovering early signaling events in plants.

Playing Cool - Winter Weather's Influence on Location-Based Gaming

Location-based games are highly dependent on the physical environment. One important but often overlooked factor is weather. In this paper we focus on winter weather, as this is a common weather that is often not designed for specifically. By performing a qualitative observation, interview, and questionnaire study of families during and after playing a GPS-based mobile game in a winter setting, this paper studies how winter conditions affected the gaming experience. Three main factors were observed to have a big impact on the gaming experience: snow, ice and cold. We outline ways these weather conditions were found to be both obstacles, and adding value to the game. Finally, we suggest design implications for winter weather, mainly; the need for short games due to cold, adaption of gameplay for movement in snow, to avoid interactions based on handling the phone, and to adapt maps to the effects of snow and ice. By explaining how winter conditions affect the gaming experience, location-based games can be better adapted for these weather conditions, and thereby help in making better design decisions.

Looks can be deceiving: contrasting temperature characteristics of two morphologically similar kelp species co-occurring in the Arctic

Two morphologically similar digitate kelp species, Laminaria digitata and Hedophyllum nigripes, co-occur along a shallow sublittoral depth gradient in the Arctic but, in contrast to L. digitata, very few ecophysiological data exist for H. nigripes. We investigated growth, survival, photosynthetic characteristics and carbon:nitrogen ratios of juvenile sporophytes, and recruitment and survival of gametophytes in genetically verified Arctic isolates of both species along temperature gradients (0–25 °C) over 14 days. Laminaria digitata gametophytes survived 23–24 °C, while sporophytes survived 21–22 °C. Hedophyllum nigripes had lower temperature affinities. Gametophytes survived 19–21 °C, while sporophytes survived 18 °C. Male gametophytes were more heat-tolerant than female gametophytes in both species. The pronounced cold adaption of H. nigripes compared to L. digitata also became apparent in different sporophyte growth optima (L. digitata: 15 °C; H. nigripes: 10 °C) and gametogenesis optima (L. digitata: 5–15 °C; H. nigripes: 0–10 °C). Higher carbon:nitrogen ratios in H. nigripes suggest an adaptation to nutrient poor Arctic conditions. The overall temperature performance of H. nigripes possibly restricts the species to Arctic–Sub-Arctic regions, while Arctic L. digitata behaved similarly to cold-temperate populations. Our data suggest that a future increase in seawater temperatures may hamper the success of H. nigripes and favour L. digitata in Arctic environments.

M2 macrophage-secreted Slit3 intensifies sympathetic nerve function in adipose tissue and enhances thermogenesis: A long-term cold adaption way

Beiging of white adipose tissue (WAT) is capable of adaptive thermogenesis and dissipating energy. The beiging processes have been associated with the increase of anti-inflammatory M2 macrophages, however the function of M2 macrophage on beiging and the underlying mechanism are not fully understood. Here we identified a macrophage cytokine Slit3 by analyzing the transcriptome of M2 macrophages collected with FACS in inguinal WAT (iWAT) of mice after cold exposure. Once released from macrophages, Slit3 bound to the ROBO1 receptor on sympathetic neuron and activated tyrosine hydrolase (TH) through PKA and CaMKⅡ signaling, and thus stimulated norepinephrine (NE) synthesis and release. NE acts on adipocytes and stimulate thermogenesis. Adoptive transfer of Slit3-overexpressing M2 macrophages to iWAT depot acquired local adipocytes with beiging phenotype and enhanced thermogenesis. In addition, mice bearing the myeloid inactivation of Slit3 were cold intolerant and gained more weight due to the lowered metabolic rate. Collectively, we demonstrate Slit3 is a macrophage cytokine and promotes beiging and thermogenesis through intensifying the sympathetic nerve function. As the expanded M2 macrophages are integral cell population in adipose tissue, the macrophage-Slit3-sympathetic neuron-adipocyte axis assures the long-term cold adaption.

Structure and dynamics of a cold-active esterase reveals water entropy and active site accessibility as the likely drivers for cold-adaptation

Cold-active esterases hold great potential for undertaking useful biotransformations at low temperatures. Here, we determined the structure of a cold active family IV esterase (EstN7) cloned from Bacillus cohnii strain N1, which has an apparent melting temperature of 26°C. EstN7 is a dimer with a classical α/β hydrolase fold. It has an acidic surface that is thought to play a role in cold-adaption by retaining solvation under changed water solvent entropy at lower temperatures. However, dynamics do not appear to play a major role in cold adaption. Comparison of B-factors with the closest related mesophilic and thermophilic esterases suggests there is little difference in dynamics with the catalytically important N-terminal cap comprising the main dynamic element. Molecular dynamics, rigidity analysis, normal mode analysis and geometric simulations of motion confirm the flexibility of the cap region but suggest that the rest of the protein is largely rigid. Rigidity analysis indicates the distribution of hydrophobic tethers is appropriate to colder conditions, where the hydrophobic effect is weaker than in mesophilic conditions due to reduced water entropy. The conformation of the cap region is significantly different to EstN7’s closest relatives, forming a bridge-like structure with reduced helical content providing more than one access tunnel through to the active site. Thus, it is likely that increased substrate accessibility and tolerance to changes in water entropy are the main drivers of EstN7’s cold adaptation rather than changes in dynamics.

Cold-Active β-Galactosidases: Insight into Cold Adaption Mechanisms and Biotechnological Exploitation

β-galactosidases (EC 3.2.1.23) catalyze the hydrolysis of β-galactosidic bonds in oligosaccharides and, under certain conditions, transfer a sugar moiety from a glycosyl donor to an acceptor. Cold-active β-galactosidases are identified in microorganisms endemic to permanently low-temperature environments. While mesophilic β-galactosidases are broadly studied and employed for biotechnological purposes, the cold-active enzymes are still scarcely explored, although they may prove very useful in biotechnological processes at low temperature. This review covers several issues related to cold-active β-galactosidases, including their classification, structure and molecular mechanisms of cold adaptation. Moreover, their applications are discussed, focusing on the production of lactose-free dairy products as well as on the valorization of cheese whey and the synthesis of glycosyl building blocks for the food, cosmetic and pharmaceutical industries.

Comparative physiology of five tropical montane songbirds reveals differential seasonal acclimatisation and cold adaption

The physiology of tropical birds is poorly understood, particularly in how it relates to local climate and changes between seasons. This is particularly true of tropical montane species, which may have sensitive thermal tolerances to local microclimates. We studied metabolic rates (using open flow respirometry), body mass and haemoglobin concentrations of five sedentary Mesoamerican songbirds between the summer and winter at two elevations (1550 m and 1950 m, respectively). We asked whether there were uniform seasonal shifts in physiological traits across species, and whether higher elevation species displayed evidence for cold tolerance. Seasonal shifts in metabolic rates differed between the three species for which data were collected. Basal metabolic rates in one species – black-headed nightingalethrushes Catharus mexicanus – were up-regulated in summer (~19% increase of winter metabolism), however two other species displayed no seasonal regulation. No species exhibited shifts in haemoglobin concentrations across season or across elevation, whereas body mass in two species was significantly higher in the summer. One species restricted to higher elevations – ruddy-capped nightingale-thrushes C. frantzii – displayed physiological traits indicative of cold-tolerance. Although only summer data were available for this species (C. frantzii), metabolic rates were constant across temperatures tested (5-34°C) and haemoglobin concentrations were significantly higher compared to the other four species. Our results suggest that seasonal acclimatisation in physiological traits is variable between species and appear unrelated to changes in local climate. As such, the distinct physiological traits observed in ruddy-capped nightingale-thrushes likely relate to historic isolation and conserved physiological traits rather than contemporary climatic adaption.