Cloning, sequencing, characterisation and implications for vaccine design of the novel dihydrolipoyl acetyltransferase of Neisseria meningitidis

Journal of Medical Microbiology
D A Ala' AldeenS P Borriello

Abstract

A lambdaZap-II expression library of Neisseria meningitidis was screened with a rabbit polyclonal antiserum (R-70) raised against c. 70-kDa proteins purified from outer membrane vesicles by elution from preparative SDS-polyacrylamide gels. Selected clones were isolated, further purified, and their recombinant pBluescript SKII plasmids were excised. The cloned DNA insert was sequenced from positive clones and analysed. Four open reading frames (ORFs) were identified, three of which showed a high degree of homology with the pyruvate dehydrogenase (E1p), dihydrolipoyl acetyltransferase (E2p) and dihydrolipoyl dehydrogenase (E3) components of the pyruvate dehydrogenase complex (PDHC) of a number of prokaryotic and eukaryotic species. Sequence analysis indicated that the meningococcal E2p (Men-E2p) contains two N-terminal lipoyl domains, an E1/E3 binding domain and a catalytic domain. The domains are separated by hinge regions rich in alanine, proline and charged residues. Another lipoyl domain with high sequence similarity to the Men-E2p lipoyl domain was found at the N-terminal of the E3 component. A further ORF, coding for a 16.5-kDa protein, was found between the ORFs encoding the E2p and E3 components. The identity and function...Continue Reading

Citations

Jul 10, 1998·Biochimica Et Biophysica Acta·A de KokA H Westphal
Feb 10, 2016·PloS One·Daniel O ConnorMarkus von Nickisch-Rosenegk
Apr 4, 2017·Pathogens and Disease·Myron Christodoulides, John Heckels
Dec 5, 2006·Molecular Microbiology·Kate L SeibMichael P Jennings
Oct 16, 1999·European Journal of Biochemistry·A F HengeveldA de Kok
Dec 2, 2005·Vaccine·D F SchuijffelP J M Nuijten

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