OGT Binding Peptide-Tagged Strategy Increases Protein O-GlcNAcylation Level in E. coli.

O-GlcNAcylation is a single glycosylation of GlcNAc mediated by OGT, which regulates the function of substrate proteins and is closely related to many diseases. However, a large number of O-GlcNAc-modified target proteins are costly, inefficient, and complicated to prepare. In this study, an OGT binding peptide (OBP)-tagged strategy for improving the proportion of O-GlcNAc modification was established successfully in E. coli. OBP (P1, P2, or P3) was fused with target protein Tau as tagged Tau. Tau or tagged Tau was co-constructed with OGT into a vector expressed in E. coli. Compared with Tau, the O-GlcNAc level of P1Tau and TauP1 increased 4~6-fold. Moreover, the P1Tau and TauP1 increased the O-GlcNAc-modified homogeneity. The high O-GlcNAcylation on P1Tau resulted in a significantly slower aggregation rate than Tau in vitro. This strategy was also used successfully to increase the O-GlcNAc level of c-Myc and H2B. These results indicated that the OBP-tagged strategy was a successful approach to improve the O-GlcNAcylation of a target protein for further functional research.

Variations in fuel use in the flight muscles of wing-dimorphic Gryllus firmus and implications for morph-specific dispersal.

Although a considerable amount of information is available on tradeoffs in wing-polymorphic insects, only limited data are available on the relationship between flight and biochemical variation within species. In the current study, we compared the biochemical basis in the dorsolongitudinal flight muscle of the wing-dimorphic sand cricket, Gryllus firmus Scudder, with respect to tradeoffs in energy resources related to morph-specific flight, including glycogen, trehalose, and triglycerides. Our results show that levels of glycogen and trehalose in long-winged adults (LW[f]) were significantly higher before dispersal, on days 5 and 7. Considering that this is the period during which long-winged adults are flight-capable, these results suggest that both glycogen and trehalose are important to flight. However, levels of triglycerides in short-winged crickets (SW) were higher than in long-winged crickets, suggesting that triglycerides are not directly related to initial flight. In SW adults, triglyceride content on days 5 and 7 was significantly higher just before lights off than at the same time on day 1 or at any other time after lights on all other days. This suggests that triglycerides are probably related to reproductive behaviors, such as mating and oviposition, in the SW morph. In addition, flight muscle water content was significantly lower in the LW(f) morph than in the SW morph.