Cell walls of Micrococcus lysodeikticus. Fractionation of the nondialyzable components from a lysozyme digest of cell walls

Imanaga, Yujiro; Park, James T.

doi:10.1021/bi00771a026

Biochemistry

1972

DOI: 10.1021/bi00771a026

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Cell walls of Micrococcus lysodeikticus. Fractionation of the nondialyzable components from a lysozyme digest of cell walls

Yujiro Imanaga¹,

James T. Park²

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Cited by 13 publications

(4 citation statements)

References 26 publications

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“…The cell walls of Micrococcus luteus consist of teichuronic acid covalently attached to peptidoglycan (4,10,12,20,25). The teichuronic acid of this bacterium is a polymer of alternating residues of D-glucose and N-acetyl-D-mannosaminuronic acid (ManNAcA), which, together, constitute a disaccharide repeat unit with the structure 4)-2-acetamido-2 -deoxy -p -D -mannopyranuronosyl -(1->6) -a-D-glucopyranosyl- (1-(9, 14).…”

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Polymer length of teichuronic acid released from cell walls of Micrococcus luteus

et al. 1990

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Teichuronic acid released from its phosphodiester linkage to peptidoglycan in the cell walls of Micrococcus luteus by mild acid treatment is resolved into a ladderlike series of bands by electrophoresis on polyacrylamide gels in the presence of borate. Each band of the ladder differs from its nearest neighbor by one disaccharide repeat unit, ----4)-2-acetamido-2-deoxy-beta-D-mannopyranuronosyl-(1----6)- alpha-D-glucopyranosyl-(1-. Acid-fragmented teichuronic acid, after conversion to the phenylamine derivative, was fractionated by preparative-scale molecular sieve column chromatography, which produced a series of elution peaks. Fast-atom-bombardment mass spectrometry of the smallest member of the series determined its molecular weight and established its identity as the phenylamine derivative of one disaccharide repeat unit of teichuronic acid. Homologous fractions of the same series were used to index the ladder of bands obtained by polyacrylamide gel electrophoresis from samples containing a more extensive distribution of polymer lengths. Nearly native teichuronic acid consists of polymers with a broad range of molecular sizes ranging from 20 to 55 disaccharide units. The most abundant species are those which have 25 to 40 repeat units. Prolonged treatment of teichuronic acid with the acid conditions used to release it from peptidoglycan causes gradual fragmentation of the teichuronic acid.

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Polymer length of teichuronic acid released from cell walls of Micrococcus luteus

et al. 1990

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“…The amount of N-acetylglucosamine detected was approximately stoichiometric with the amount of phosphate in the complex. Partial fragmentation of teichuronic acid provides an explanation of the previous erroneous identification of the reducing end residue.Micrococcus luteus cell walls consist of a peptidoglycan matrix to which teichuronic acid is covalently attached (2,8,12,13,29). Although the structure of the disaccharide repeat unit of teichuronic acid has been determined by chemical procedures (9) and confirmed by 13C nuclear magnetic resonance (NMR) spectroscopy (15), the precise nature of the linkage of teichuronic acid to peptidoglycan has remained in dispute.…”

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confidence: 99%

“…Micrococcus luteus cell walls consist of a peptidoglycan matrix to which teichuronic acid is covalently attached (2,8,12,13,29). Although the structure of the disaccharide repeat unit of teichuronic acid has been determined by chemical procedures (9) and confirmed by 13C nuclear magnetic resonance (NMR) spectroscopy (15), the precise nature of the linkage of teichuronic acid to peptidoglycan has remained in dispute.…”

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confidence: 99%

Teichuronic acid reducing terminal N-acetylglucosamine residue linked by phosphodiester to peptidoglycan of Micrococcus luteus

et al. 1990

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Teichuronic acid-peptidoglycan complex isolated from Micrococcus luteus cells by lysozyme digestion in osmotically stabilized medium was treated with mild acid to cleave the linkage joining teichuronic acid to peptidoglycan. This labile linkage was shown to be the phosphodiester which joins N-acetylglucosamine, the residue located at the reducing end of the teichuronic acid, through its anomeric hydroxyl group to a 6-phosphomuramic acid, a residue of the glycan strand of peptidoglycan. 31P nuclear magnetic resonance spectroscopy of the lysozyme digest of cell walls demonstrated the presence of a phosphodiester which was converted to a phosphomonoester by the conditions which released teichuronic acid from cell walls. Reduction of acid-liberated reducing end groups by NaB3H4 followed by complete acid hydrolysis yielded [3H] glucosaminitol from the true reducing end residue of teichuronic acid and [3H]glucitol from the sites of fragmentation of teichuronic acid. The amount of N-acetylglucosamine detected was approximately stoichiometric with the amount of phosphate in the complex. Partial fragmentation of teichuronic acid provides an explanation of the previous erroneous identification of the reducing end residue.Micrococcus luteus cell walls consist of a peptidoglycan matrix to which teichuronic acid is covalently attached (2,8,12,13,29). Although the structure of the disaccharide repeat unit of teichuronic acid has been determined by chemical procedures (9) and confirmed by 13C nuclear magnetic resonance (NMR) spectroscopy (15), the precise nature of the linkage of teichuronic acid to peptidoglycan has remained in dispute. A small amount of phosphate is present in M. luteus cell walls (2), and much of it can be recovered from strong acid hydrolysates as muramic acid-6-phosphate (24). Glucosamine-6-phosphate has also been detected (18,27). A phosphodiester has been suggested as part of the covalent link between peptidoglycan and teichuronic acid in M. luteus (10,25,26) just as it serves to link teichoic acids and other cell wall polymers to peptidoglycan in other microorganisms (4,16,17,19,23).Nasir-ud-Din and Jeanloz (25) claimed that glucose is the reducing end residue of teichuronic acid which links to peptidoglycan through the phosphodiester. Later Nasirud-Din et al. (26) reported experimental results which purported to show that glucose is the reducing end residue of teichuronic acid. Evidence presented by Hase and Matsushima (10) indicated instead that the reducing end residue is N-acetylglucosamine. Our studies of in vitro teichuronic acid biosynthesis (14, 31) showed that biosynthesis depends on the initial step which incorporated N-acetylglucosamine-1-phosphate into a carrier lipid-activated intermediate. Furthermore, the N-acetylglucosamine residue remains in product teichuronic acid at the reducing end, suggesting that teichuronic acid in native cell walls has an N-acetylglucosamine residue at its potentially reducing terminal residue.We report here that the true reducing end residue of teichuronic ...

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“…The cell wall of Micrococcus luteus (lysodeikticus) consists of peptidoglycan and a teichuronic acid that contains glucose and N-acetylmannosaminuronic acid (ManNAcUA) (21). The teichuronic acid is a polysaccharide with repeating units of -4--ManNAcUJA--L'kGlc-e-'-- (6) which is covalently linked to peptidoglycan (4,10,17). The linkage includes a phosphodiester between position 6 of muramic acid (14) or glucosamine (18) residues of peptidoglycan and position 1 of the N-acetylglucosamine (GlcNAc) residue at the reducing end of the teichuronic acid (7).…”

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Elongation of teichuronic acid chains by a wall-membrane preparation from Micrococcus luteus

Traxler¹,

Goustin²,

Anderson³

1982

J Bacteriol

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A wall-plus-membrane preparation from Micrococcus luteus catalyzes the incorporation of [14C]glucose from UDP-[14C]glucose, into two fractions of teichuronic acid, which is the cell wall polysaccharide consisting of alternating residues of glucose and N-acetylmannosaminuronic acid (ManNAcUA). Membrane-associated teichuronic acid was extracted from the wall-membrane fraction of reaction mixtures by sodium dodecyl sulfate. The synthesis of membraneassociated teichuronic acid required UDP-glucose, UDP-ManNAcUA, and UDP-N-acetylglucosamine and was inhibited by tunicamycin. Glucose incorporated

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Cell walls of Micrococcus lysodeikticus. Fractionation of the nondialyzable components from a lysozyme digest of cell walls

Cited by 13 publications

References 26 publications

Polymer length of teichuronic acid released from cell walls of Micrococcus luteus

Polymer length of teichuronic acid released from cell walls of Micrococcus luteus

Teichuronic acid reducing terminal N-acetylglucosamine residue linked by phosphodiester to peptidoglycan of Micrococcus luteus

Elongation of teichuronic acid chains by a wall-membrane preparation from Micrococcus luteus

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