BiP (Immunoglobulin Binding Protein) is a member of the Hsp70 chaperones that participates in protein folding in the endoplasmic reticulum. The function of BiP relies on cycles of ATP hydrolysis driving the binding and release of its substrate proteins. It still remains unknown how BiP affects the protein folding pathway and there has been no direct demonstration showing which folding state of the substrate protein is bound by BiP, as previous work has used only peptides. Here, we employ optical tweezers for single molecule force spectroscopy experiments to investigate how BiP affects the folding mechanism of a complete protein and how this effect depends on nucleotides. Using the protein MJ0366 as the substrate for BiP, we performed pulling and relaxing cycles at constant velocity to unfold and refold the substrate. In the absence of BiP, MJ0366 unfolded and refolded in every cycle. However, when BiP was added, the frequency of folding events of MJ0366 significantly decreased, and the loss of folding always occurred after a successful unfolding event. This process was dependent on ATP and ADP, since when either ATP was decreased or ADP was added, the duration of periods without folding events increased. Our results show that the affinity of BiP for the substrate protein increased in these conditions, which correlates with previous studies in bulk. Therefore, we conclude that BiP binds to the unfolded state of MJ0366 and prevents its refolding, and that this effect is dependent on both the type and concentration of nucleotides.

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Abstract:The zooplankton assemblages in southern Chilean Patagonia are characterized by calanoid dominance and low species number that is observable under oligotrophic status and wide conductivity values, whereas at mesotrophic status the daphnids are dominant with high species number, and finally at hyper-saline environments halophilic species such as Artemia persimilis and/or the calanoid Boeckella poopensis predominate. In the present study data of different lakes and ponds between 45-53• S were analyzed, with the aim to determine potential structures at different sites. For this purpose a null model based in guild structure was applied, considering each guild a different kind of water body (lake, small lake, permanent pond, ephemeral pond, and saline lake). The results revealed in two simulations that guild are structured. These results are similar with other descriptions on the basis of null models that revealed a random pattern of species associations for similar ecosystems due to many species repeated in all or practically all studied sites or similarities of ecological features. Ecological and biogeographical topics were discussed.