Structural evidence of <b>α</b>‐aminoacylated lipoproteins of <i>Staphylococcus aureus</i>

Asanuma, Miwako; Kurokawa, Kenji; Ichikawa, Rie; Ryu, Kyoung-Hwa; Chae, Jun-Ho; Dohmae, Naoshi; Lee, Bok Luel; Nakayama, Hiroshi

doi:10.1111/j.1742-4658.2010.07990.x

Cited by 48 publications

(64 citation statements)

References 33 publications

(65 reference statements)

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“…Fragments of tryptic lipopeptides observed in MALDI MS/MS spectra are mainly C-terminal-containing sequence ions (y-type), because the positive charge tends to localize at C-terminal lysine or arginine residues of the peptides. We confirmed that both diacyl and triacyl synthetic peptides [Pam2 (dipalmitoyl)-CSK 4 and Pam3-CSK 4 ] could be analyzed with respect to organic solvent extraction and ionization (3). The efficiencies of ionization of the synthetic diacyl and triacyl synthetic peptides are slightly different: the triacyl peptide was slightly more easily ionized than its diacyl counterpart in our MALDI-TOF MS analysis.…”

Section: Methodssupporting

confidence: 63%

“…Proteins separated by SDS-PAGE were subjected to ingel digestion with trypsin, and the resulting digests (10 to 20 l) were acidified with 1 l of 70% formic acid and then mixed with the same volume of a chloroform/methanol mixture (2/1 [vol/vol]) to extract lipidmodified peptides. Finally, matrix-assisted laser desorption ionizationtime of flight (MALDI-TOF) mass spectrometry (MS) was performed on the lipopeptides in the lower organic phase as previously described (3).…”

Section: Methodsmentioning

confidence: 99%

“…The E. coli-type Lnt is conserved in most Gram-negative bacteria and in high-GC-content, Gram-positive bacteria, such as Mycobacterium tuberculosis and Streptomyces coelicolor, which were shown to contain triacyl lipoproteins (36,38). However, in the absence of the E. coli-type Lnt homolog, some low-GC-content, Gram-positive bacteria and mycoplasmas, such as Staphylococcus aureus, Mycoplasma pneumoniae, and Acholeplasma laidlawii, contain the N-acylated triacyl lipoprotein form, too (3,18,31). Furthermore, we recently discovered new classes of N-acylated lipid-modified structures in low-GC-content Gram-positive bacteria, the lyso form and the N-acetyl form (18).…”

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confidence: 99%

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Environment-Mediated Accumulation of Diacyl Lipoproteins over Their Triacyl Counterparts in Staphylococcus aureus

et al. 2012

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Bacterial lipoproteins are believed to exist in only one specific lipid-modified structure, such as the diacyl form or the triacyl form, in each bacterium. In the case of Staphylococcus aureus , recent extensive matrix-assisted laser desorption ionization–time of flight (MALDI-TOF) mass spectrometry analysis revealed that S. aureus lipoproteins exist in the α-aminoacylated triacyl form. Here, we discovered conditions that induce the accumulation of diacyl lipoproteins that lack α-aminoacylation in S. aureus . The accumulation of diacyl lipoproteins required a combination of conditions, including acidic pH and a post-logarithmic-growth phase. High temperatures and high salt concentrations additively accelerated the accumulation of the diacyl lipoprotein form. Following a post-logarithmic-growth phase where S. aureus MW2 cells were grown at pH 6, SitC lipoprotein was found almost exclusively in its diacyl structure rather than in its triacyl structure. This is the first report showing that the environment mediates lipid-modified structural alterations of bacterial lipoproteins.

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Section: Methodssupporting

confidence: 63%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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Environment-Mediated Accumulation of Diacyl Lipoproteins over Their Triacyl Counterparts in Staphylococcus aureus

et al. 2012

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“…While our work was under review, an important study by Asanuma et al (27) appeared where the authors report that SitC and four other major lipoproteins from Staphylococcus aureus are triacylated despite the apparent absence of the lnt (N-acyltransferase) gene in this organism. It therefore appears that triacylation of lipoproteins is a global phenomenon in the bacterial world.…”

Section: Discussionmentioning

confidence: 99%

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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“…The identification of the protein band was performed as described (45). The protein band was excised from the gel, destained with 50% (vol/vol) methanol, and dried by vacuum centrifugation.…”

Section: Generation Of Complemented Strainsmentioning

confidence: 99%

Polyester synthesis genes associated with stress resistance are involved in an insect–bacterium symbiosis

Kim

Won

Nikoh

et al. 2013

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

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Significance This study reports a previously unrecognized involvement of polyhydroxyalkanoate (PHA), known as a bacterial endocellular storage polymer, in an insect–bacterium symbiosis. Many bacteria in the environment accumulate PHA granules within their cells, which provide resistance to nutritional depletion and other environmental stresses. Here we demonstrate that synthesis and accumulation of PHA in the symbiont cells are required for normal symbiotic association with, and, consequently, positive fitness effects for the host insect. The requirement of PHA for symbiosis suggests that, contrary to the general expectation, the within-host environment may be, at least in some aspects, stressful for the symbiotic bacteria.

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Structural evidence of α‐aminoacylated lipoproteins of Staphylococcus aureus

Cited by 48 publications

References 33 publications

Environment-Mediated Accumulation of Diacyl Lipoproteins over Their Triacyl Counterparts in Staphylococcus aureus

Environment-Mediated Accumulation of Diacyl Lipoproteins over Their Triacyl Counterparts in Staphylococcus aureus

The Acylation State of Surface Lipoproteins of Mollicute Acholeplasma laidlawii

Polyester synthesis genes associated with stress resistance are involved in an insect–bacterium symbiosis

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