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

2019 ◽  
Author(s):  
Shijun Bao ◽  
Xiaoqin Ding ◽  
Shengqing Yu ◽  
Chan Ding
Keyword(s):  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Surface Membrane ◽  
Binding Activity ◽  
Expression Vectors ◽  
Economic Losses ◽  
Beta Subunits ◽  
Related Factors ◽  
Human Fibronectin ◽  
Dehydrogenase Complex

ABSTRACTMycoplasma 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 (pdhAandpdhB, respectively) of MS WVU1853 strain in GenBank were used to amplify thepdhAandpdhBgenes of MS WVU1853 strain through PCR. Then the prokaryotic expression vectors pET-pdhA and pET-pdhB were constructed and were expressed inEscherichia coliBL21(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 were distributed both on the surface membrane and within soluble cytosolic fractions of MS cells. The rMSPDHA and rMSPDHB presented binding activity with chicken Plg and human Fn. The rabbit anti-rMSPDHA and anti-rMSPDHB sera had distinct mycoplasmacidal efficacy in the presence of guinea pig complement, and the adherence of MS to DF-1 cells pretreated with Plg was effectively inhibited by treatment with anti-rMSPDHA or anti-rMSPDHB sera. Hence, the study indicates that the surface-associated MSPDHA and MSPDHB are the adhesion-related factors of MS that contributes to bind to Plg/Fn and adhesion to DF-1 cells.

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

2021 ◽  
pp. 104851
Author(s):  
Shijun Bao ◽  
Xiaoqin Ding ◽  
Shengqing Yu ◽  
Xiaoyong Xing ◽  
Chan Ding
Keyword(s):  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Beta Subunits ◽  
Mycoplasma Synoviae ◽  
Dehydrogenase Complex

scholarly journals Biosynthesis, import and processing of precursor polypeptides of mammalian mitochondrial pyruvate dehydrogenase complex

1988 ◽  
Vol 251 (3) ◽  
pp. 817-823 ◽  
Author(s):  
O G L De Marcucci ◽  
G M Gibb ◽  
J Dick ◽  
J G Lindsay
Keyword(s):  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Beta Subunits ◽  
Intact Cells ◽  
Carbonyl Cyanide ◽  
Rat Liver Cells ◽  
Immune Precipitation ◽  
Additional Segment ◽  
Kinetics Of ◽  
Dehydrogenase Complex

An immunological analysis has been conducted of early events in the biosynthesis, import and assembly of the mammalian pyruvate dehydrogenase complex (PDC). For this purpose, monospecific polyclonal antisera were produced against the intact assembly from ox heart, Mr 8.5 x 10(6), and each of its component polypeptides, E1 alpha, E1 beta, E2, E3 and protein X. Optimal detergent-based incubation mixtures were developed for obtaining clean immunoprecipitation of PDC polypeptides and their precursors from [35S]methionine-labelled extracts of PK-15 (pig kidney), NBL-1 (bovine kidney) and BRL (Buffalo Rat liver) cells. In PK-15 cells, independent higher Mr species, corresponding to precursors of the E2, E1 alpha and E1 beta subunits of PDC, could be detected by immune precipitation and fluorography after incubation of intact cells for 4 h with [35S]methionine and 1-2 mM-2,4-dinitrophenol or 10-15 microM-carbonyl cyanide p-trifluoromethoxyphenylhydrazone. Similar precursor states could be observed in uncoupler-treated BRL or NBL-1 cells. Pre-E1 alpha, pre-E1 beta and also pre-E3, have signal sequences in the Mr range 1500-3000 while pre-E2 contains a long additional segment of Mr 7000-9000. All of these forms exhibit similar kinetics of processing to the mature subunits with a transit time of 10-12 min. In NBL-1 cells, E3 is present in the immune complexes formed with anti-PDC serum whereas this is not the case in PK-15 cells. Thus, there are significant variations in the affinity of lipoamide dehydrogenase (E3) for the E2 core structure in different species. Pre-E1 alpha accumulates only poorly in PK-15 cells and is aberrantly processed on removal of uncoupler. This precursor is markedly more stable in NBL-1 and BRL cells. The lack of detection of a precursor form of component X is also discussed.

The Pyruvate Dehydrogenase Complex as a Target for Gene Therapy

2003 ◽  
Vol 3 (3) ◽  
pp. 239-245 ◽  
Author(s):  
Peter Stacpoole ◽  
Renius Owen ◽  
Terence Flotte
Keyword(s):  
Gene Therapy ◽  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Dehydrogenase Complex

scholarly journals Amidination of pyruvate dehydrogenase complex of Escherichia coli under denaturing conditions

1979 ◽  
Vol 177 (1) ◽  
pp. 136-137 ◽  
Author(s):  
G Hale ◽  
E A Hooper ◽  
R N Perham
Keyword(s):  
Escherichia Coli ◽  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Dehydrogenase Complex

scholarly journals A Resolution of the Escherichia coli Pyruvate Dehydrogenase Complex

1961 ◽  
Vol 236 (4) ◽  
pp. 1013-1018 ◽  
Author(s):  
Anne D. Gounaris ◽  
Lowell P. Hager
Keyword(s):  
Escherichia Coli ◽  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Dehydrogenase Complex
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