Sterols and Acylated Proteins in Mycoplasmas

Rottem, Shlomo

doi:10.1006/bbrc.2001.2023

Cited by 11 publications

(9 citation statements)

References 42 publications

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“…For several proteins, MALDI/ MS-MS analyses yielded a more precise acylation pattern because the N-acylated peptide mass allowed for the determination of the modification pattern. It has been suggested earlier the Mollicutes are characterized by diacylation (45). However, we determined that A. laidlawii has three acyl residues (50).…”

Section: General Genomic Featuresmentioning

confidence: 72%

Complete Genome and Proteome of Acholeplasma laidlawii

et al. 2011

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We present the complete genome sequence and proteogenomic map for Acholeplasma laidlawii PG-8A (class Mollicutes, order Acholeplasmatales, family Acholeplasmataceae). The genome of A. laidlawii is represented by a single 1,496,992-bp circular chromosome with an average G؉C content of 31 mol%. This is the longest genome among the Mollicutes with a known nucleotide sequence. It contains genes of polymerase type I, SOS response, and signal transduction systems, as well as RNA regulatory elements, riboswitches, and T boxes. This demonstrates a significant capability for the regulation of gene expression and mutagenic response to stress. Acholeplasma laidlawii and phytoplasmas are the only Mollicutes known to use the universal genetic code, in which UGA is a stop codon. Within the Mollicutes group, only the sterol-nonrequiring Acholeplasma has the capacity to synthesize saturated fatty acids de novo. Proteomic data were used in the primary annotation of the genome, validating expression of many predicted proteins. We also detected posttranslational modifications of A. laidlawii proteins: phosphorylation and acylation. Seventy-four candidate phosphorylated proteins were found: 16 candidates are proteins unique to A. laidlawii, and 11 of them are surface-anchored or integral membrane proteins, which implies the presence of active signaling pathways. Among 20 acylated proteins, 14 contained palmitic chains, and six contained stearic chains. No residue of linoleic or oleic acid was observed. Acylated proteins were components of mainly sugar and inorganic ion transport systems and were surface-anchored proteins with unknown functions.

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Section: General Genomic Featuresmentioning

confidence: 72%

Complete Genome and Proteome of Acholeplasma laidlawii

et al. 2011

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“…This incorporation of cholesterol obtained from their hosts into glycolipids appears to be a special feature of Borrelia sp., shared only with a few other pathogenic bacteria like Mycoplasma sp. (29,30) or Helicobacter sp. (31).…”

Section: Discussionmentioning

confidence: 99%

Acylated Cholesteryl Galactosides Are Specific Antigens of Borrelia Causing Lyme Disease and Frequently Induce Antibodies in Late Stages of Disease

Stübs

Fingerle

Wilske

et al. 2009

Journal of Biological Chemistry

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“…Free cholesterol or cholesterol esters are components of several bacterial membranes such as Mycoplasma (20), Helicobacter pylori (21), Micrococcus lysodeikticus, Bacillus megaterium, and Proteus mirabilis (22). Cholesteryl glycosides have been identified in Mycoplasmas (20,23), H. pylori (21), and Borrelia hermsi (24), in all of which, the carbohydrate was glucose.…”

Section: Discussionmentioning

confidence: 99%

“…Cholesteryl glycosides have been identified in Mycoplasmas (20,23), H. pylori (21), and Borrelia hermsi (24), in all of which, the carbohydrate was glucose. Cholesteryl Galactoside is now identified in bacteria.…”

Section: Discussionmentioning

confidence: 99%

A newly discovered cholesteryl galactoside from Borrelia burgdorferi

Ben-Menachem

Kübler-Kiełb²,

Coxon³

et al. 2003

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

150

128

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Two major glycolipids, which comprise Ϸ36% of the total lipid mass from Borrelia burgdorferi, the etiological agent of Lyme disease, were investigated. We determined the fatty acid type, sugar identity, anomeric configuration, and substituent type and position. The structures were identified as cholesteryl 6-O-acyl-␤-D-galactopyranoside (B. burgdorferi glycolipid 1, BbGL-I), and 1,2-di-O-acyl-3-O-␣-D-galactopyranosyl-sn-glycerol (BbGL-II). The major fatty acids were palmitate and oleate. The structures were corroborated by gas-liquid chromatography MS, matrix-assisted laser desorption͞ionization time-of-flight spectroscopy, fast atom bombardment MS, detailed NMR spectrometry, and metabolic labeling. This is a previously undescribed demonstration of a cholesteryl galactoside in bacteria. Lipopolysaccharide was not detected in B. burgdorferi. The two glycolipids have several properties suggesting they may function as lipopolysaccharide: both are main components of the bacterial membrane, surface exposed, and have a three-domain structure. BbGL-I elicited specific antibodies in mice and rabbits, and BbGL-II elicited antibodies that reacted with both glycolipids.T he etiological agent of Lyme disease, Borrelia burgdorferi, is transmitted to humans through the bite of Ixodes ticks. Lyme disease is a multisystem infection, which affects the skin, joints, nervous system, and heart (1). The licensed vaccine (LYMErix, SmithKline Beecham) contains recombinant lipidated OspA. Although lipopolysaccharide (LPS) has been identified in several spirochaetales, such as Leptospira (2) and Treponema (3), there is no evidence for the presence of LPS in Borrelia species (4).Two surface-exposed glycolipids identified in B. burgdorferi react with sera from Lyme disease patients (5-7). These immunoreactive glycolipids have been characterized as monogalactosyl diacylglycerolipids (8). It has been proposed that these glycolipids differ only in their fatty acid composition. We designated these glycolipids as B. burgdorferi glycolipid I (BbGL-I) and B. burgdorferi glycolipid II (BbGL-II). We show that although BbGL-II is a monogalactosyl diacylglycerol as reported (8), BbGL-I has the unique structure cholesteryl 6-O-acyl-␤-D-galactopyranoside. Experimental ProceduresOrganism and Growth Conditions. B. burgdorferi strains B31 (ATCC 35210), BL303 (courtesy of G. Wormser, Division of Infectious Diseases, New York Medical College, Valhalla) and N40 (courtesy of L. Bockenstedt, Department of Internal Medicine, Yale University School of Medicine, New Haven, CT) were cultivated in BSK-H medium (Sigma). Media were inoculated with 2% (vol͞vol) of a frozen culture and incubated statically at 37°C for 72 h to the mid-exponential growth phase. Cells were harvested by centrifugation at 12,000 ϫ g for 30 min, washed three times with cold PBS, and stored at Ϫ20°C.Lipid Extraction and Purification. Lipids were extracted from washed cells as described (9). The chloroform was removed, and the dried lipids (0.1-0.2 mg͞mg cell) dissolved in 1-2 ml of chlorofor...

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Sterols and Acylated Proteins in Mycoplasmas

Cited by 11 publications

References 42 publications

Complete Genome and Proteome of Acholeplasma laidlawii

Complete Genome and Proteome of Acholeplasma laidlawii

Acylated Cholesteryl Galactosides Are Specific Antigens of Borrelia Causing Lyme Disease and Frequently Induce Antibodies in Late Stages of Disease

A newly discovered cholesteryl galactoside from Borrelia burgdorferi

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