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

Abstract

Mycoplasma synoviae (MS) is an important pathogen, causing enormous economic losses to the poultry industry worldwide every year. Therefore, the studies on MS will lay the foundation for diagnosis, prevention and treatment of MS infection. In this study, primers designed based on the sequences of pyruvate dehydrogenase complex (PDC) E1 alpha and beta subunit genes (pdhA and pdhB, respectively) of MS WVU1853 strain in GenBank were used to amplify the pdhA and pdhB genes of MS WVU1853 strain through PCR. Then the prokaryotic expression vectors pET-pdhA and pET-pdhB were constructed and were expressed in Escherichia coli BL21(DE3) cells. Subsequently, the recombinant proteins rMSPDHA and rMSPDHB were purified and anti-rMSPDHA and anti-rMSPDHB sera were prepared by immunizing rabbits, respectively. Finally, the subcellular localization of PDHA and PDHB in MS, binding activity of rMSPDHA and rMSPDHB to chicken plasminogen (Plg) and human fibronectin (Fn), complement-dependent mycoplasmacidal assays, and adherence and adherence inhibition assays were accomplished. The results showed that PDHA and PDHB .

Characterization and Evaluation of a Novel Conserved Membrane Antigen P35 of Mycoplasma synoviae

“…MS is distributed worldwide and causes serious economic losses to the global poultry industry every year. Research on MS can lay the foundation for the prevention, diagnosis, and treatment of MS-related diseases ( 20 ). Previous studies have screened several immunogenic proteins from MS isolates ( 21 ).…”

“…The development of more effective and cheaper vaccines requires to identify and characterize antigens of MS. Also, investigation on pathogenicity mechanism and serological detection methods are hampered by the lack of known and well-characterized MS antigens. However, only a few antigens have been identified so far, such as lipoprotein MSPB, elongation factor EF-Tu, enolase, NADH oxidase, hemagglutinin MSPA, ATP synthase beta chain, trigger factor, pyruvate kinase, chaperone DnaK, and pyruvate dehydrogenase complex E1 alpha and beta subunits ( 20 , 21 ). But the immunogenicity and antigenicity of these proteins have not been thoroughly studied.…”

The immunogenicity identification and adherence characterization analysis of acetate kinase of Mycoplasma synoviae

“…For the distribution of ACK in MS cells, ELISA was processed as previously described simultaneously [20].…”

“…In the Mycoplasma sp., there are some enzymes involved in metabolism have been found to be associated with pathogenicity, such as fructose-1,6bisphosphate aldolase [15], NADH oxidase [16], Glyceraldehyde-3-phosphate dehydrogenase [17], methylenetetrahydrofolate-tRNA-(uracil-5-)-methyltransferase [18], triosephosphate isomerase [19] and so on. In our previous studies, several enzymatic proteins of MS antigen and adhesion factor have been identi ed, including α-and β-subunits of pyruvate decarboxylase [20] and Enolase [21]. To enhance the understanding of the pathogenic mechanism of MS, the immunogenicity and the adherence characteristic of MS ACK were further analyzed in this study.…”

Mycoplasma synoviae LP78 is a fibronectin/plasminogen binding protein, putative adhesion, and potential diagnostic antigen

“…The second stage entails the correctly binding of the microorganism-specific ligands to the relevant host cell receptors, primarily via the surface proteins ( Razin and Jacobs, 1992 ). Variable lipoprotein hemagglutinin (vlhA), enolase, pyruvate dehydrogenase alpha (PdhA) and beta (PdhB) subunits, dihydrolipoamide dehydrogenase (PdhD), NADH oxidase and other surface proteins from M. synoviae have been identified to be involved in adhesion ( Noormohammadi et al, 1997 ; Bao et al, 2014 , 2021 ; Hu et al, 2022 ; Qi et al, 2022 ). Additionally, most microorganisms bind to a range of host extracellular matrix (ECM) components, such as fibronectin (Fn), collagen, elastin, and laminin via interactions between microbial adhesins and host cell receptors ( Chagnot et al, 2012 ).…”