Fastloc-GPS reveals daytime departure and arrival during long-distance migration and the use of different resting strategies in sea turtles

Determining the time of day that animals initiate and end migration, as well as variation in diel movement patterns during migration, provides insights into the types of strategy used to maximise energy efficiency and ensure successful completion of migration. However, obtaining this level of detail has been difficult for long-distance migratory marine species. Thus, we investigated whether the large volume of highly accurate locations obtained by Argos-linked Fastloc-GPS transmitters could be used to identify the time of day that adult green (n = 8 turtles, 9487 locations) and loggerhead (n = 46 turtles, 47,588 locations) sea turtles initiate and end migration, along with potential resting strategies during migration. We found that departure from and arrival at breeding, stopover and foraging sites consistently occurred during the daytime, which is consistent with previous findings suggesting that turtles might use solar visual cues for orientation. Only seven turtles made stopovers (of up to 6 days and all located close to the start or end of migration) during migration, possibly to rest and/or refuel; however, observations of day versus night speed of travel indicated that turtles might use other mechanisms to rest. For instance, turtles travelled 31% slower at night compared to day during their oceanic crossings. Furthermore, within the first 24 h of entering waters shallower than 100 m towards the end of migration, some individuals travelled 72% slower at night, repeating this behaviour intermittently (each time for a one-night duration at 3–6 day intervals) until reaching the foraging grounds. Thus, access to data-rich, highly accurate Argos-linked Fastloc-GPS provided information about differences in day versus night activity at different stages in migration, allowing us, for the first time, to compare the strategies used by a marine vertebrate with terrestrial land-based and flying species

Application of virus-like particles (VLP) to NMR characterization of viral membrane protein interactions

The membrane proteins of viruses play critical roles in the virus life cycle and are attractive targets for therapeutic intervention. Virus Like Particles (VLP) present the possibility to study the biochemical and biophysical properties of viral membrane proteins in their native environment. Specifically, the VLP constructs contain the entire protein sequence and are comprised of native membrane components including lipids, cholesterol, carbohydrates and cellular proteins. In this study we prepare VLP containing full-length hemagglutinin (HA) or neuraminidase (NA) from influenza and characterize their interactions with small molecule inhibitors. Using HA-VLP, we first show that VLP samples prepared using the standard sucrose gradient purification scheme contain significant amounts of serum proteins, which exhibit high potential for non-specific interactions, thereby complicating NMR studies of ligand-target interactions. We then show that the serum contaminants may be largely removed with the addition of a gel filtration chromatography step. Next, using HA-VLP we demonstrate that WaterLOGSY (WL) NMR is significantly more sensitive than Saturation Transfer Difference (STD) NMR for the study of ligand interactions with membrane bound targets. In addition, we compare the ligand orientation to HA embedded in VLP with that of recombinant HA by STD NMR. In a subsequent step, using NA-VLP we characterize the kinetic and binding properties of substrate analogs and inhibitors of NA, including study of the H274Y-NA mutant, which leads to wide spread resistance to current influenza antivirals. In summary, our work suggests that VLP have high potential to become standard tools in biochemical and biophysical studies of viral membrane proteins, particularly when VLP are highly purified and combined with control VLP containing native membrane proteins