Selective Acylation of Plasma Membrane Proteins of Mycoplasma mycoides subsp. mycoides SC, the Contagious Bovine Pleuropneumonia Agent

Jan, G.; Fontenelle, Catherine; Verrier, Florence; Hénaff, Michel Le; Wróblewski, Henri

doi:10.1007/s002849900007

Cited by 19 publications

(16 citation statements)

References 16 publications

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“…The existence of A. laidlawii lipoproteins in triacylated form is unexpected because a gene encoding recognizable homologs of N-acyltransferases responsible for incorporation of the third fatty acid in lipoproteins from Gram-negative bacteria is absent from the A. laidlawii genome. Identification of triacylation as the major (perhaps the only) state of A. laidlawii lipoproteins is in contrast with published data, which suggest the predominant involvement of O-ester-linked fatty acids but not amide-linked fatty acids in protein acylation in A. laidlawii (14,26). In the latter work, the authors showed that globomycin, a cyclic lipopeptide specifically inhibiting signal peptidase II, had no dramatic effect on the extent of acylation of A. laidlawii lipoproteins, suggesting the preferential absence of amide-linked acyl chains.…”

Section: Discussioncontrasting

confidence: 54%

“…In the latter work, the authors showed that globomycin, a cyclic lipopeptide specifically inhibiting signal peptidase II, had no dramatic effect on the extent of acylation of A. laidlawii lipoproteins, suggesting the preferential absence of amide-linked acyl chains. The reasons for the discrepancy with previous data are unknown but may be related to the fact that both Le Hénaff and co-workers (14) and Nyström et al (26) analyzed fatty acids that were released from lipoproteins by various chemical treatments. The failure to detach even trace amounts of non-covalently associated lipids from their lipoprotein preparations or imperfection of releasing reactions could have heavily influenced the outcome of their measurements.…”

Section: Discussioncontrasting

confidence: 48%

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The Acylation State of Surface Lipoproteins of Mollicute Acholeplasma laidlawii

Serebryakova

Demina

Galyamina

et al. 2011

Journal of Biological Chemistry

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Acylation of the N-terminal Cys residue is an essential, ubiquitous, and uniquely bacterial posttranslational modification that allows anchoring of proteins to the lipid membrane. In Gram-negative bacteria, acylation proceeds through three sequential steps requiring lipoprotein diacylglyceryltransferase, lipoprotein signal peptidase, and finally lipoprotein N-acyltransferase. The apparent lack of genes coding for recognizable homologs of lipoprotein N-acyltransferase in Gram-positive bacteria and Mollicutes suggests that the final step of the protein acylation process may be absent in these organisms. In this work, we monitored the acylation state of eight major lipoproteins of the mollicute Acholeplasma laidlawii using a combination of standard two-dimensional gel electrophoresis protein separation, blotting to nitrocellulose membranes, and MALDI-MS identification of modified N-terminal tryptic peptides. We show that for each A. laidlawii lipoprotein studied a third fatty acid in an amide linkage on the N-terminal Cys residue is present, whereas diacylated species were not detected. The result thus proves that A. laidlawii encodes a lipoprotein N-acyltransferase activity. We hypothesize that N-acyltransferases encoded by genes non-homologous to N-acyltransferases of Gram-negative bacteria are also present in other mollicutes and Gram-positive bacteria.Essential cellular activities such as transport, sensing, signal transduction, growth, adhesion, and pathogenesis require proteins localized on the cell surface. One of the strategies to anchor a protein at the lipid membrane is covalent modification of proteins by fatty acids or other lipids. All bacteria rely on acylation of N-terminal Cys residues of lipoproteins to allow tight anchorage on the membrane surface (1). This uniquely bacterial modification was first observed in 1969 in a major outer membrane protein of Escherichia coli called Braun's lipoprotein and later detected in every bacterial organism studied (2). Adhesion proteins and substrate-binding subunits of ATPbinding cassette (ABC) 2 transporters are known to be attached to the membrane in this manner. Thousands of proteins predicted to be lipoproteins "by similarity" according to the presence of consensus sequences of Sec Type II or Tat Type II signal peptides are also thought to be acylated (3).Bacteria of the Mollicutes class are widespread in nature and are characterized by the lack of a rigid cell wall and drastically reduced genome sizes. In fact, representatives of this class are the simplest self-replicating organisms able to independently subsist, generate energy, and adapt to changing environments (4 and references therein). Most mollicutes depend on cholesterol and are partially or totally incapable of fatty acid synthesis and therefore depend on the host or the culture medium for a constant supply of these substances (5). Acholeplasma laidlawii is a convenient model mollicute because of its relatively simple nutritional requirements and fast growth rate. A. laidlawii can synthesize limit...

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Section: Discussioncontrasting

confidence: 54%

Section: Discussioncontrasting

confidence: 48%

The Acylation State of Surface Lipoproteins of Mollicute Acholeplasma laidlawii

Serebryakova

Demina

Galyamina

et al. 2011

Journal of Biological Chemistry

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“…The N-acyltransferase gene responsible for N acylation has not been detected in the M. pneumoniae, M. genitalium, and M. penetrans genomes (11,14,39) either. However, a study of the ratio of N-amide and O-ester bonds in M. gallisepticum and M. mycoides suggested that such mycoplasmas possess diacylated and triacylated lipoproteins (18). Furthermore, the resistance to Edman degradation of proteins from M. mycoides also indicates the presence of N acylation (5).…”

Section: Discussionmentioning

confidence: 99%

A Triacylated Lipoprotein fromMycoplasma genitaliumActivates NF-κB through Toll-Like Receptor 1 (TLR1) and TLR2

2008

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Mycoplasma genitalium is a sexually transmitted bacterial pathogen that causes nongonococcal chlamydianegative urethritis, mucopurulent cervicitis, endometritis, pelvic inflammatory disease, and tubal factor infertility in humans. However, pathogenic agents that induce inflammatory responses have not been identified in M. genitalium. In this study, we examined the involvement of Toll-like receptors ( Mycoplasma genitalium is an important, emerging sexually transmitted bacterial pathogen that is capable of eliciting a wide range of reproductive tract disease syndromes (3,19,20,28). M. genitalium has been identified as a causative agent of nongonococcal, chlamydia-negative urethritis (8, 50, 51) in men and of mucopurulent cervicitis, endometritis, pelvic inflammatory disease, and tubal factor infertility in women (6,7,24,33,34,45). However, pathogenic agents that induce an inflammatory response, such as endotoxin and exotoxin, have not been identified in M. genitalium.Recently, it has been reported that Toll-like receptors (TLRs) with a pattern recognition receptor function play a critical role in early innate recognition and in the inflammatory responses by the host defense against invading microbes (1, 21). Of 10 TLR family members reported, TLR2, TLR4, TLR5, and TLR9 have been implicated in the recognition of different bacterial components. Peptidoglycan, lipoarabinomannan, zymosan, and lipoproteins from various microorganisms are recognized by TLR2 (2,4,22,26,46,47,49,52). On the other hand, lipopolysaccharide, bacterial flagellin, and bacterial DNA are recognized by TLR4, TLR5, and TLR9, respectively (12,13,16,35). These TLR family members have been shown to activate NF-B via interleukin-1R-associated signal molecules, including myeloid differentiation protein (MyD88), interleukin-1R-activated kinase, tumor necrosis factor receptor-associated factor 6, and NF-B-inducing kinase (27).In this study, we examined the involvement of TLRs in activation of the immune response by lipoproteins from M. genitalium and their active components responsible for NF-B activation. MG149, a triacylated lipoprotein, was found to activate NF-B through TLR1 and TLR2. MATERIALS AND METHODSCells. Cells of a human kidney cell line, 293T, were cultured in Dulbecco modified Eagle medium containing 10% fetal calf serum, 2 mM L-glutamine, 100 U/ml penicillin G, and 100 g/ml streptomycin. To exclude the possibility of mycoplasma infections in the cell cultures, the cell suspensions were inoculated onto pleuropneumonialike organism agar medium once a month. To check the contamination of mycoplasma, cells were also stained with 4Ј,6-diamidino-2-phenylindole (DAPI) once a week. Synthesis of lipopeptide. Lipopeptide was synthesized by Bio-Synthesis (Lewisville, TX).TX-114 phase partitioning. M. genitalium G37 was cultured in SP-4 medium to the beginning of the stationary phase and then pelleted by centrifugation for 10 min at 12,000 ϫ g. M. genitalium cells were inactivated by incubation at 60°C for 30 min. Triton X-114 (TX-114) phase partitio...

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“…An N-acyltransferase gene responsible for N-acylation has not been detected in the M. pneumoniae, Mycoplasma genitalium, Mycoplasma penetrans and U. parvum genomes (Fraser et al, 1995;Glass et al, 2000;Himmelreich et al, 1996;Sasaki et al, 2002) either. However, a study of the ratio of N-amide and Oester bonds in M. gallisepticum and Mycoplasma mycoides suggests that such mycoplasmas possess diacylated and triacylated lipoproteins (Jan et al, 1996b). Furthermore, the resistance to Edman degradation of proteins from M. mycoides also indicates the presence of N-acylation (Chambaud et al, 1999).…”

Section: Discussionmentioning

confidence: 99%

Ureaplasma parvum lipoproteins, including MB antigen, activate NF-κB through TLR1, TLR2 and TLR6

2008

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Ureaplasma species (Ureaplasma parvum and Ureaplasma urealyticum) are commonly isolated pathogens from the female reproductive tract and are associated with perinatal diseases in humans. Inappropriate induction of inflammatory responses may be involved in the occurrence of such diseases; however, pathogenic agents that induce the inflammatory response have not been identified in ureaplasmas. In this study, we examined the involvement of Toll-like receptors (TLRs) in the activation of the immune response by U. parvum lipoproteins, as well as the U. parvum components responsible for nuclear factor kB (NF-kB) activation. The Triton X-114 (TX-114) detergent phase of U. parvum was found to induce NF-kB through TLR2. The active components of the TX-114 detergent phase were lipoproteins, such as multiple banded (MB) antigen, UU012 and UU016 of U. parvum. The activation of NF-kB by these lipoproteins was inhibited by dominant negative (DN) constructs of TLR1 and DN TLR6. Thus, the lipoproteins from U. parvum were found to activate NF-kB through TLR1, TLR2 and TLR6. Furthermore, these lipoproteins possessed an ability to induce tumour necrosis factor-alpha (TNF-a) in mouse peritoneal macrophages. INTRODUCTIONUreaplasmas are wall-less bacteria belonging to the Mycoplasmataceae.Although Ureaplasma species (Ureaplasma parvum and Ureaplasma urealyticum) are common commensals of the urogenital tract of humans, ureaplasmas have frequently been recognized as important pathogens associated with non-gonococcal urethritis (Shepard, 1954), chorioamnionitis (Cassell et al., 1986;Maher et al., 1994), preterm birth (Jelsema, 2006;Kataoka et al., 2006; Ollikainen et al., 1998), infertility (TaylorRobinson, 1986;Xu et al., 1997), pneumonia in neonates (Waites et al., 1989) and bronchopulmonary dysplasia in neonates (Schelonka et al., 2005). The mechanisms by which ureaplasmas cause such diseases are unclear, but the inappropriate induction of inflammatory responses may be involved. It has been reported that the levels of interleukin-6 (IL-6), tumour necrosis factor-alpha (TNF-a), IL-1b and IL-8 in intra-amniotic fluid infected with ureaplasmas are increased (Yoon et al., 1998). Moreover, macrophages are capable of producing such cytokines and nitric oxide in response to U. urealyticum (Li et al., 2000a, b). However, pathogenic agents such as endotoxin and exotoxin that induce an inflammatory response have not been identified in ureaplasmas.It has been widely known that Toll-like receptors (TLRs) with the function of pattern-recognition receptors play critical roles in early innate recognition and inflammatory responses by the host against invading microbes (Akira & Takeda, 2004;Kopp & Medzhitov, 1999). Among 10 TLR family members reported, TLR2, TLR4, TLR5 and TLR9 have been implicated in the recognition of different bacterial components. Peptidoglycan (PGN), lipoarabinomannan, zymosan and lipoproteins from various microorganisms are recognized by TLR2 (Aliprantis et al., 1999;Brightbill et al., 1999;Lien et al., 1999;Means et al., 1999...

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Selective Acylation of Plasma Membrane Proteins of Mycoplasma mycoides subsp. mycoides SC, the Contagious Bovine Pleuropneumonia Agent

Cited by 19 publications

References 16 publications

The Acylation State of Surface Lipoproteins of Mollicute Acholeplasma laidlawii

The Acylation State of Surface Lipoproteins of Mollicute Acholeplasma laidlawii

A Triacylated Lipoprotein fromMycoplasma genitaliumActivates NF-κB through Toll-Like Receptor 1 (TLR1) and TLR2

Ureaplasma parvum lipoproteins, including MB antigen, activate NF-κB through TLR1, TLR2 and TLR6

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