Ng‐MIP, a surface‐exposed lipoprotein of Neisseria gonorrhoeae, has a peptidyl‐prolyl cis/trans isomerase (PPIase) activity and is involved in persistence in macrophages

Leuzzi, Rosanna; Serino, Laura; Scarselli, María; Savino, Silvana; Fontana, Maria Rita; Monaci, Elisabetta; Taddei, Annarita; Fischer, Gunter; Rappuoli, Rino; Pizza, Mariagrazia

doi:10.1111/j.1365-2958.2005.04859.x

Cited by 113 publications

(116 citation statements)

References 27 publications

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“…PA4370 encodes an insulin-cleaving metalloproteinase (IcmP) whose activity was detected in outer membrane protein fractions; however, these authors did not report the presence of a predicted lipoprotein signal peptide (3). PA3262 encodes a probable peptidyl-prolyl cis-trans isomerase with homology to MIP (macrophage infectivity potentiator), which is a surface-exposed, outer membrane lipoprotein in Neisseria and Chlamydia (9,14). Regions encoding the native promoters and entire open reading frames (lacking stop codons) of predicted lipoproteins PA4370 and PA3262 were cloned upstream of mCherry.…”

Section: Resultsmentioning

confidence: 99%

“…Three fatty acids are attached to the N-terminal cysteine residue that immediately follows the cleavage site of lipoprotein signal peptides (13). The lipids integrate into the periplasm-facing leaflet of either membrane, leaving most lipoproteins exposed to the periplasm (13), although some lipoproteins are surface exposed (9,16). We have previously used fluorescent microscopy to visualize red fluorescent lipoproteins (lipoRFPs) directly in both the outer and inner membranes of Escherichia coli and other Enterobacteriaceae (10).…”

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confidence: 99%

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Novel Inner Membrane Retention Signals inPseudomonas aeruginosaLipoproteins

2008

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The ultimate membrane localization and function of most of the 185 predicted Pseudomonas aeruginosa PAO1 lipoproteins remain unknown. We constructed a fluorescent lipoprotein, CSFP OmlA -ChFP, by fusing the signal peptide and the first four amino acids of the P. aeruginosa outer membrane lipoprotein OmlA to the monomeric red fluorescent protein mCherry (ChFP). When cells were plasmolyzed with 0.5 M NaCl, the inner membrane separated from the outer membrane and formed plasmolysis bays. This permits the direct observation of fluorescence in either the outer or inner membrane. CSFP OmlA -ChFP was shown to localize in the outer membrane by fluorescence microscopy and immunoblotting analysis of inner and outer membrane fractions. The site-directed substitution of the amino acids at positions ؉2, ؉3, and ؉4 in CSFP OmlA -ChFP was performed to test the effects on lipoprotein localization of a series of amino acid sequences selected from a panel of predicted lipoproteins. We confirmed Asp ؉2 and Lys ؉3 Ser ؉4 function as inner membrane retention signals and identified four novel inner membrane retention signals:, and CQ ؉2 G ؉3 S ؉4 . These inner membrane retention signals are found in 5% of the 185 predicted P. aeruginosa lipoproteins. Full-length chimeras of predicted lipoproteins PA4370 and PA3262 fused to mCherry were shown to reside in the inner membrane and showed a nonuniform or patchy distribution in the membrane. The optical sectioning of cells producing PA4370 CGDD -ChFP and PA3262 CDSQ -ChFP by confocal microscopy improved the resolution and indicated a helix-like localization pattern in the inner membrane. The method described here permits the in situ visualization of lipoprotein localization and should work equally well for other membrane-associated proteins.

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Section: Resultsmentioning

confidence: 99%

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confidence: 99%

Novel Inner Membrane Retention Signals inPseudomonas aeruginosaLipoproteins

2008

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“…The remaining glycoproteins include a peptidyl prolyl-isomerase (Ng1225), 2 solute-binding proteins associated with ABC transport systems (Ng0372 and Ng1494), and 2 lipoproteins of unknown function (Ng1043 and Ng2139). We further note that with the exceptions of PilE as well as Ng1225 and Ng2139 [for which evidence for surface exposure exists (24,25)], the glycoproteins are predicted to function in the periplasmic compartment, although their precise sites of localization await confirmation.…”

Section: N a A S G T A S A Pmentioning

confidence: 99%

Broad spectrum O-linked protein glycosylation in the human pathogen Neisseria gonorrhoeae

Vik

Aas

Anonsen

et al. 2009

Proc. Natl. Acad. Sci. U.S.A.

146

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Protein glycosylation is an important element of biologic systems because of its significant effects on protein properties and functions. Although prominent within all domains of life, O-linked glycosylation systems modifying serine and threonine residues within bacteria and eukaryotes differ substantially in target protein selectivity. In particular, well-characterized bacterial systems have been invariably dedicated to modification of individual proteins or related subsets thereof. Here we characterize a general O-linked glycosylation system that targets structurally and functionally diverse groups of membrane-associated proteins in the Gram-negative bacterium Neisseria gonorrhoeae, the etiologic agent of the human disease gonorrhea. The 11 glycoproteins identified here are implicated in activities as varied as protein folding, disulfide bond formation, and solute uptake, as well as both aerobic and anaerobic respiration. Along with their common trafficking within the periplasmic compartment, the protein substrates share quasi-related domains bearing signatures of low complexity that were demonstrated to encompass sites of glycan occupancy. Thus, as in eukaryotes, the broad scope of this system is dictated by the relaxed specificity of the glycan transferase as well as the bulk properties and context of the proteintargeting signal rather than by a strict amino acid consensus sequence. Together, these findings reveal previously unrecognized commonalities linking O-linked protein glycosylation in distantly related life forms.bacteria ͉ glycoprotein ͉ pilin ͉ post-translational modification ͉ pgl

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“…Proteomic studies carried out in our laboratory identified the high abundance of a 29-kDa meningococcal MIP protein (the product of gene NMB1567, NEIS1487) in the OM (20). It has been also reported that the gonococcal homologue Neisseria gonorrhoeae macrophage infectivity potentiator (Ng-MIP) was a surface-exposed lipoprotein in N. gonorrhoeae (21). We showed in a previous study (22) that MIP is highly conserved, and in a collection of well-characterized meningococcal isolates (differing in serogroup, serotype, and serosubtype), isolated from carriers or patients, we found only three distinct MIP sequence types (designated I, II, and III).…”

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confidence: 99%

Recombinant Protein Truncation Strategy for Inducing Bactericidal Antibodies to the Macrophage Infectivity Potentiator Protein of Neisseria meningitidis and Circumventing Potential Cross-Reactivity with Human FK506-Binding Proteins

et al. 2015

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A recombinant macrophage infectivity potentiator (rMIP) protein of Neisseria meningitidis induces significant serum bactericidal antibody production in mice and is a candidate meningococcal vaccine antigen. However, bioinformatics analysis of MIP showed some amino acid sequence similarity to human FK506-binding proteins (FKBPs) in residues 166 to 252 located in the globular domain of the protein. To circumvent the potential concern over generating antibodies that could recognize human proteins, we immunized mice with recombinant truncated type I rMIP proteins that lacked the globular domain and the signal leader peptide (LP) signal sequence (amino acids 1 to 22) and contained the His purification tag at either the N or C terminus (C-term). The immunogenicity of truncated rMIP proteins was compared to that of full (i.e., full-length) rMIP proteins (containing the globular domain) with either an N-or C-terminal His tag and with or without the LP sequence. By comparing the functional murine antibody responses to these various constructs, we determined that C-term His truncated rMIP (؊LP) delivered in liposomes induced high levels of antibodies that bound to the surface of wild-type but not ⌬mip mutant meningococci and showed bactericidal activity against homologous type I MIP (median titers of 128 to 256) and heterologous type II and III (median titers of 256 to 512) strains, thereby providing at least 82% serogroup B strain coverage. In contrast, in constructs lacking the LP, placement of the His tag at the N terminus appeared to abrogate bactericidal activity. The strategy used in this study would obviate any potential concerns regarding the use of MIP antigens for inclusion in bacterial vaccines. N eisseria meningitidis (meningococcus) infections contribute significantly to mortality and morbidity worldwide (1). Implementation of capsular polysaccharide-protein conjugate vaccines against serogroups A, C, Y, and W into the routine immunization schedules of developed countries has been successful (2-5), but this approach cannot be used for serogroup B strains. The polysaccharide capsule of serogroup B meningococci (MenB) shows structural mimicry of human fetal brain neural cell adhesion molecules (6). Licensed MenB vaccines based on lipooligosaccharide (LOS)-depleted outer membrane (OM) vesicles (V) have been used to control serosubtype strain-specific clonal outbreaks of MenB infection, e.g., in Norway (7), Cuba (8), Brazil (9), and New Zealand (10), but they do not provide cross-strain protection (11). Recently, the 4CMenB (Bexsero) vaccine, developed using a genome-based reverse-vaccinology approach (12), has received a license from the European Union and has been recommended by the Joint Committee for Vaccination and Immunization for the routine vaccination of infants in the United Kingdom since 2014. The vaccine consists of the factor H binding protein (fHbp, fused to GNA2091 carrier protein), neisserial heparin binding protein (NHBA, fused to GNA1030 carrier protein), and an adhesin, NadA, mixed with the Men...

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Ng‐MIP, a surface‐exposed lipoprotein of Neisseria gonorrhoeae, has a peptidyl‐prolyl cis/trans isomerase (PPIase) activity and is involved in persistence in macrophages

Cited by 113 publications

References 27 publications

Novel Inner Membrane Retention Signals inPseudomonas aeruginosaLipoproteins

Novel Inner Membrane Retention Signals inPseudomonas aeruginosaLipoproteins

Broad spectrum O-linked protein glycosylation in the human pathogen Neisseria gonorrhoeae

Recombinant Protein Truncation Strategy for Inducing Bactericidal Antibodies to the Macrophage Infectivity Potentiator Protein of Neisseria meningitidis and Circumventing Potential Cross-Reactivity with Human FK506-Binding Proteins

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