Mycoplasma synoviaeenolase is a plasminogen/fibronectin binding protein

Abstract

BackgroundMycoplasma synoviae is an avian pathogen that can lead to respiratory tract infections and arthritis in chickens and turkeys, resulting in serious economic losses to the poultry industry. Enolase reportedly plays important roles in several bacterial pathogens, but its role in M. synoviae has not been established. Therefore, in this study, the enolase encoding gene (eno) of M. synoviae was amplified from strain WVU1853 and expressed in E. coli BL21 cells. Then the enzymatic activity, immunogenicity and binding activity with chicken plasminogen (Plg) and human fibronectin (Fn) was evaluated.ResultsWe demonstrated that the recombinant M. synoviae enolase protein (rMsEno) can catalyze the conversion of 2-phosphoglycerate (2-PGA) to phosphoenolpyruvate (PEP), the Km and Vmax values of rMsEno were 1.1 × 10−3 M and 0.739 μmol/L/min, respectively. Western blot and immuno-electron microscopy analyses confirmed that enolase was distributed on the surface and within the cytoplasm of M. synoviae cells. The binding assays demonstrated that rMsEno was able to bind to chicken Plg and human Fn proteins. A complement-dependent mycoplasmacidal assay demonstrated that rabbit anti–rMsEno serum had distinct mycoplasmacidal efficacy in the presence of complement, which also confirmed that enolase was distributed on the surface of M. synoviae. An inhibition assay showed that the adherence of M. synoviae to DF-1 cells pre-treated with Plg could be effectively inhibited by treatment with rabbit anti-rMsEno serum.ConclusionThese results reveal that M. synoviae enolase has good catalytic activity for conversion of 2-PGA to PEP, and binding activity with chicken Plg and human Fn. Rabbit anti–rMsEno serum displayed an obvious complement-dependent mycoplasmacidal effect and adherent inhibition effect. These results suggested that the M. synoviae enolase plays an important role in M. synoviae metabolism, and could potentially impact M. synoviae infection and immunity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12917-014-0223-6) contains supplementary material, which is available to authorized users.

Network of Surface-Displayed Glycolytic Enzymes in Mycoplasma pneumoniae and Their Interactions with Human Plasminogen

“…Surface-displayed glycolytic enzymes have also been demonstrated in mycoplasmas such as the important veterinary species Mycoplasma bovis (17), Mycoplasma gallisepticum (18), Mycoplasma suis (19,20), and Mycoplasma synoviae (21), as well in the human pathogens Mycoplasma fermentans (22) and Mycoplasma genitalium (23). As described for other bacteria species, enolase (Eno) and glutaraldehyde-6-phosphate dehydrogenase (GapA) were confirmed as the main surface-located glycolytic enzymes.…”

Expression, purification, and biological characterization of <i>Anaplasma phagocytophilum</i> enolase

“…Interestingly, rApEno demonstrated the highest enzymatic activity at 50°C and pH 8.0 (Figure 3). Moreover, rApEno presented high K m and V max values during 2-PGA to PEP conversion, when compared to other infectious intracellular pathogens (13,20). This suggests that ApEno is highly adaptable to a changing surrounding environment.…”

Characterization of pyruvate dehydrogenase complex E1 alpha and beta subunits ofMycoplasma synoviae

“…Complement-dependent 147 mycoplasmacidal activity of rabbit anti-rMSPDHA or anti-rMSPDHB sera was determined as 148 described previously (Bao et al, 2014). MS, grown to mid-logarithmic phase, was washed three 149 times with PBS by centrifugation at 5,000×g for 10 min at 4°C, and re-suspended in PBS at the final concentration of 6×10 3 CFU/mL.…”