Propeptide processing and proteolytic activity of proenzymes of the Staphylococcal and Enterococcal GluV8-family protease.

Proenzymes with various lengths of propeptides have been observed in GluV8 from Staphylococcus aureus and GluSE from S. epidermidis. However, the production mechanism of these proenzymes and roles of truncated propeptides have yet to be elucidated. Here we demonstrate that shortening of propeptide commonly occurs in an auto-catalytic manner in GluV8-family members, including those from coagulase negative Staphylococci and Enterococcus faecalis. Accompanied with propeptide shortening, the pro-mature junction (Asn/Ser-1-Val1) becomes more susceptible towards the hetero-catalytic maturation enzymes. The auto-catalytic propeptide truncation is not observed in Ser169Ala inert molecules of GluV8-family members. A faint proteolytic activity of proenzymes from Staphylococcus caprae and E. faecalis is detected. In addition, proteolytic activity of proenzyme of GluV8 carrying Arg-3AlaAsn-1 is demonstrated with synthetic peptide substrates LLE/Q-MCA. These results suggest that GluV8-family proenzymes with shortened propeptides intrinsically possess proteolytic activity and are involved in the propeptide shortening that facilitates the final hetero-catalytic maturation.

Immunologic reactivity on one year follow-up of subjects without allergy to Hymenoptera stings.

We studied Hymenoptera stings in 72 pest-control operators without any previous systemic reactions to Hymenoptera stings, and investigated their venom-specific IgE levels in serial specimens collected over one year. At the initial evaluation, venom-specific IgE was present in 25 (34.7%) of 72 pest-control operators, and venom-specific IgE titer significantly decreased as the time interval from the last sting increased (p < 0.001). In most cases, venom-specific IgE disappeared less than 3 years after the last sting. On the other hand, the ratio of subjects with positive CAP for venom-specific IgE was significantly increased with an elevation of total serum IgE level (p < 0.001). After the one year follow-up, venom-specific IgE titer in the 25 subjects with positive CAP decreased significantly (p = 0.026). Total serum IgE level modified the decline significantly (p = 0.011), but the time interval from the last sting did not. In elevated total IgE level (>250 IU/ml), the decline of venom-specific IgE tended to be slow.