Detection of Specific Lipids in Mycobacteria by Infrared Spectroscopy

Smith, Donald W.; Randali, H. M.; Maclennan, A. P.; Putney, R. K.; Rao, Srivatsa V.

doi:10.1128/jb.79.2.217-229.1960

Cited by 47 publications

(23 citation statements)

References 22 publications

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“…In some cases, the 6-deoxytalose moiety may be further extended with haptenic oligosaccharides resulting in highly antigenic serovar specific GPLs (Kolk et al ., 1989;Daffe and Lemmassu, 2000). They have been studied from a structural point of view during the last 50 years (Smith et al ., 1960a;Barrow and Brennan, 1982; for review Billman-Jacobe, 2004), but the first molecular structure of a GPL was solved in the late 60s (Smith et al ., 1960b). The synthesis of GPLs by organic chemistry has only been recently achieved (Heidelberg and Martin, 2004).…”

Section: Introductionmentioning

confidence: 99%

Gap, a mycobacterial specific integral membrane protein, is required for glycolipid transport to the cell surface

Sondén¹,

Kocı́ncová²,

Deshayes³

et al. 2005

Molecular Microbiology

112

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SummaryThe cell envelope of mycobacteria is a complex multilaminar structure that protects the cell from stresses encountered in the environment, and plays an important role against the bactericidal activity of immune system cells. The outermost layer of the mycobacterial envelope typically contains species-specific glycolipids. Depending on the mycobacterial species, the major glycolipid localized at the surface can be either a phenolglycolipid or a peptidoglycolipid (GPL). Currently, the mechanism of how these glycolipids are addressed to the cell surface is not understood. In this study, by using a transposon library of Mycobacterium smegmatis and a simple dye assay, six genes involved in GPLs synthesis have been characterized. All of these genes are clustered in a single genomic region of approximately 60 kb. We show by biochemical analyses that two non-ribosomal peptide synthetases, a polyketide synthase, a methyltransferase and a member of the MmpL family are required for the biosynthesis of the GPLs backbone. Furthermore, we demonstrate that a small integral membrane protein of 272 amino acids named Gap ( gap : GPL addressing protein) is specifically required for the transport of the GPLs to the cell surface. This protein is predicted to contain six transmembrane segments and possesses homologues across the mycobacterial genus, thus delineating a new protein family. This Gap family represents a new paradigm for the transport of small molecules across the mycobacterial envelope, a critical determinant of mycobacterial virulence.

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Section: Introductionmentioning

confidence: 99%

Gap, a mycobacterial specific integral membrane protein, is required for glycolipid transport to the cell surface

Sondén¹,

Kocı́ncová²,

Deshayes³

et al. 2005

Molecular Microbiology

112

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“…Chemical differentiation has been an attractive possibility since the demonstration by Anderson of the uniqueness and complexity of tubercle cell wall lipids (1). As early as 1954, Smith and associates characterized mycobacterial species by using column chromatography and infrared spectroscopy to detect species specific myococerenates of phthioceral (28)(29)(30). Later, several studies used thin-layer chromatographic analysis of lipids and glycolipids to classify many types of nontuberculous mycobacteria (4,9,10,(14)(15)(16)31).…”

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confidence: 99%

Identification of Clinical Isolates of Mycobacteria with Gas-Liquid Chromatography Alone

1979

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Identification of 18 mycobacterial species was performed by analysis of profiles obtained by using gas-liquid chromatography. Organisms were saponified in methanolic NaOH, and the reaction mixture was treated with BF3 in methanol and extracted with a hexane-chloroform mixture. An identification scheme was developed from 128 stock strains and tested against a collection of 79 clinical isolates. By using gas-liquid chromatographic profiles alone, 58% of specimens were correctly identified to species level, and an additional 41% were correctly identified to a group of two or three organisms. Use in a clinical laboratory over a 2-month period proved chromatography to be as accurate as and more rapid than concurrent biochemical testing. Of 81 isolates tested, 64% were identified to species level by chromatography alone. An additional 35% were differentiated to the same groups of two or three organisms as found in our analysis of stock strains. These groups consisted of: Mycobacterium tuberculosis, M. bovis, and M. xenopi; M. avium complex, M. gastri, and M. scrofulaceum; or M. fortuitum and M. chelonei. Identification to species level from these groups could usually be done by colonial morphology alone and could always be done by the addition of one selected biochemical test. This study demonstrated the practical application of gas-liquid chromatography in the identification of mycobacteria in a clinical laboratory. In particular, all strains of M. gordonae and M. kansasii were identified to species level. M. tuberculosis was definitively identified in 85% of cases. When it could not be definitely identified, the only alternatives were M. bovis and M. xenopi, both of which are rare causes of infection.

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“…Mycoside D was 1 to 2 g dissolved in a few ml of h&x3ne-benzene demonstrated in the lipids from these cells, and (50:50, v/v), were added to the p"ked column, was eluted from the column by ether-methanol and the elution was begun with the same solvent (80:20, v/v). Reference to the spectra of this and~~~~~~~~~~~8:0 v/v).ue Reference to the spectrae ofithi and continued according to the sequence given column now suggests the presence of another by Smith et al (1960). Then 100 ml of each solsubstance, tentatively called component X, presvent were collected; the elution with any given ent in small amounts in several fractions eluted solvent was continued until the amount of main ether-methanol (95:5, v/v).…”

mentioning

confidence: 99%

“…The solvents were removed and infrared spectra cr oan were recorded for each fraction, according to the 1956. procedure described earlier (Smith et al, 1960).…”

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confidence: 99%

A MUTANT OF A SCOTOCHROMOGENIC MYCOBACTERIUM DETECTED BY COLONY MORPHOLOGY AND LIPID STUDIES

1962

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Detection of Specific Lipids in Mycobacteria by Infrared Spectroscopy

Cited by 47 publications

References 22 publications

Gap, a mycobacterial specific integral membrane protein, is required for glycolipid transport to the cell surface

Gap, a mycobacterial specific integral membrane protein, is required for glycolipid transport to the cell surface

Identification of Clinical Isolates of Mycobacteria with Gas-Liquid Chromatography Alone

A MUTANT OF A SCOTOCHROMOGENIC MYCOBACTERIUM DETECTED BY COLONY MORPHOLOGY AND LIPID STUDIES

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