Sequence comparison of outer membrane phospholipases A: Implications for structure and for the catalytic mechanism

Brok, Ronald G. P. M.; Boots, A.P.; Dekker, Niek; Verheij, Hubertus M.; Tommassen, Jan

doi:10.1016/s0923-2508(99)80017-5

Cited by 23 publications

(28 citation statements)

References 20 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The presence of a putative pldA gene was also demonstrated with the genome sequences of the human pathogens N. meningitidis and Neisseria gonorrhoeae (2). Interestingly, the deduced meningococcal and gonococcal OMPLA proteins differ in only a few amino acid residues.…”

mentioning

confidence: 69%

“…It was discovered and studied extensively in Escherichia coli, where it was shown to exhibit phospholipase A 1 and A 2 , lysophospholipase, and diacylglyceride lipase activity (38; for reviews, see references 8 and 40). The elucidation of many bacterial genome sequences demonstrated that OMPLA is widespread among gram-negative bacteria (2). Under normal conditions, OMPLA is present as an inactive monomer in the outer membrane.…”

mentioning

confidence: 99%

See 1 more Smart Citation

Function of Neisserial Outer Membrane Phospholipase A in Autolysis and Assessment of Its Vaccine Potential

Bos¹,

Tefsen²,

Voet

et al. 2005

Infect Immun

View full text Add to dashboard Cite

Outer membrane phospholipase A (OMPLA) is an outer membrane-localized enzyme, present in many gram-negative bacterial species. It is implicated in the virulence of several pathogens. Here, we investigated the presence, function, and vaccine potential of OMPLA in the human pathogen Neisseria meningitidis. Immunoblot analysis showed the presence of OMPLA in 28 out of 33 meningococcal strains investigated. The OMPLAnegative strains all contained a pldA gene, but these alleles contained premature stop codons. All six Neisseria gonorrhoeae strains tested, but only two out of seven commensal neisserial strains investigated, expressed OMPLA, showing that OMPLA is expressed by, but not limited to, many pathogenic neisserial strains. The function of OMPLA was investigated by assessing the phenotypes of isogenic strains, expressing no OMPLA, expressing wild-type levels of OMPLA, or overexpressing OMPLA. OMPLA exhibited phospholipase activity against endogenous phospholipids. Furthermore, OMPLA was characterized as an autolysin that acted under specific conditions, such as prolonged growth of the bacteria. The vaccine potential of the protein was investigated by immunizing mice with in vitro refolded, recombinant OMPLA. High levels of antibody titers were obtained, but the murine sera were neither bactericidal nor protective. Also, convalescent patients and vaccinee sera did not contain detectable levels of anti-OMPLA antibodies, indicating that OMPLA may not be sufficiently immunogenic to be included in a meningococcal vaccine.

show abstract

mentioning

confidence: 69%

mentioning

confidence: 99%

Function of Neisserial Outer Membrane Phospholipase A in Autolysis and Assessment of Its Vaccine Potential

Bos¹,

Tefsen²,

Voet

et al. 2005

Infect Immun

View full text Add to dashboard Cite

show abstract

“…Since haemolytic PLA 2 activity with a cytolytic effect is considered as a virulence factor in pathogenic bacteria (Songer, 1997) and is present in strains of the closely related species C. jejuni and C. coli (Brok et al, 1998), the presence of similar activity in both genomospecies of C. concisus clinical isolates supports the possible virulence role of this phospholipase/haemolysin in the pathogenicity of C. concisus. Studies have been undertaken by our team to analyse the pldA gene in C. concisus as a potential virulence factor associated with tissue destruction that may be related to intestinal inflammation as a consequence of infection by C. concisus.…”

Section: Pcr Analysis Of Pla Genementioning

confidence: 85%

“…4 kDa (Song et al, 1999), while the molecular mass for the outermembrane phospholipase A (OMPLA) monomer in Escherichia coli is 31 kDa (de Geus et al, 1983;Brok et al, 1998). It could be suggested that the molecular mass of the enzyme monomer in C. concisus is approximately 34 kDa, by comparison with the major immunoreactive band on the blot, while one of the other minor bands (67 kDa) may represent the dimeric state of the enzyme.…”

Section: Immunoblot Analysis Of Omps and Hesmentioning

confidence: 99%

Characterization of a haemolytic phospholipase A2 activity in clinical isolates of Campylobacter concisus

Istivan

Coloe

Fry

et al. 2004

Journal of Medical Microbiology

View full text Add to dashboard Cite

A membrane-bound, haemolytic phospholipase A 2 (PLA 2 ) activity was detected in clinical strains of Campylobacter concisus isolated from children with gastroenteritis. The clinical strains were assigned into two molecular groups (genomospecies) based on PCR amplification of their 23S rDNA. This calcium-dependent, heat-stable, haemolytic PLA 2 activity was detected in strains from both genomospecies. A crude haemolysin extract (CHE) was initially prepared from cellular outermembrane proteins of these isolates and was further fractionated by ultrafiltration. The haemolytic activity of the extracted fraction (R30) was retained by ultrafiltration using a 30 kDa molecular mass cut-off filter, and was designated haemolysin extract (HE). Both CHE and HE had PLA 2 activity and caused stable vacuolating and cytolytic effects on Chinese hamster ovary cells in tissue culture. Primers for the conserved region of pldA gene (phospholipase A gene) from Campylobacter coli amplified a gene region of 460 bp in all tested isolates, confirming the presence of a homologous PLA gene sequence in C. concisus. The detection of haemolytic PLA 2 activity in C. concisus indicates the presence of a potential virulence factor in this species and supports the hypothesis that C. concisus is a possible opportunistic pathogen.

show abstract

“…pylori OMPLA [35,36]. Furthermore, the Asn156Asp substitution in E. coli OMPLA yields an active species [46].…”

Section: Active Sitementioning

confidence: 99%

Bacterial phospholipase A: structure and function of an integral membrane phospholipase

Snijder

Dijkstra

2000

Biochimica et Biophysica Acta (BBA) - Molecular and Cell Biolog

View full text Add to dashboard Cite

Sequence comparison of outer membrane phospholipases A: Implications for structure and for the catalytic mechanism

Cited by 23 publications

References 20 publications

Function of Neisserial Outer Membrane Phospholipase A in Autolysis and Assessment of Its Vaccine Potential

Function of Neisserial Outer Membrane Phospholipase A in Autolysis and Assessment of Its Vaccine Potential

Characterization of a haemolytic phospholipase A2 activity in clinical isolates of Campylobacter concisus

Bacterial phospholipase A: structure and function of an integral membrane phospholipase

Contact Info

Product

Resources

About