Primary structure of cyanelle peptidoglycan of Cyanophora paradoxa: a prokaryotic cell wall as part of an organelle envelope.
The peptidoglycan layer surrounding the photosynthetic organelles (cyanelles) of the protist Cyanophora paradoxa is thought to be a relic of their cyanobacterial ancestors. The separation of muropeptides by gel filtration and reverse-phase high-performance liquid chromatography revealed four different muropeptide monomers. A number of muropeptides were identical in retention behavior to muropeptides of Escherichia coli, while others had remarkably long retention times with respect to their sizes, as indicated by gel filtration. Molecular mass determination by plasma desorption and matrix-assisted laser desorption ionization mass spectrometry showed that these unusual muropeptides had molecular masses greater by 112 Da or a multiple thereof than those of ones common to both species. Fast atom bombardment-tandem mass spectrometry of these reduced muropeptide monomers allowed the localization of the modification to D-glutamic acid. High-resolution fast atom bombardment-mass spectrometry and amino acid analysis revealed N-acetylputrescine to be the substituent (E. Pittenauer, E. R. Schmid, G. Allmaier, B. Pfanzagl, W. Löffelhardt, C. Quintela, M. A. de Pedro, and W. Stanek, Biol. Mass Spectrom. 22:524-536, 1993). In addition to the 4 monomers already known, 8 dimers, 11 trimers, and 6 tetramers were characterized. An average glycan chain length of 51 disaccharide units was determined by the transfer of [U-14C]galactose to the terminal N-acetylglucosamine residues of cyanelle peptidoglycan. The muropeptide pattern is discussed with respect to peptidoglycan biosynthesis and processing.
Pfanzagl
B
B
Institut für Biochemie und Molekulare Zellbiologie, Universität Wien, Vienna, Austria.
Zenker
A
A
Pittenauer
E
E
Allmaier
G
G
Martinez-Torrecuadrada
J
J
Schmid
E R
ER
De Pedro
M A
MA
Löffelhardt
W
W
D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002240:Carbohydrate Sequence; D002473:Cell Wall / Q000737:chemistry; D003960:Diaminopimelic Acid / Q000032:analysis; D056890:Eukaryota / Q000737:chemistry*; D008969:Molecular Sequence Data; D010457:Peptidoglycan / Q000032:analysis / Q000737:chemistry*; D018087:Plastids / Q000737:chemistry*
D000596:Amino Acids; D010457:Peptidoglycan; D003960:Diaminopimelic Acid