Tuomanen E, Schwartz J, Sande S, Light K, Gage D
Rockefeller University, New York, New York 10021.
J Biol Chem. 1989 Jul 5;264(19):11093-8.
The composition of the peptidoglycan of Bordetella pertussis and the nature of its turnover products was determined by a new combination of analytical techniques: high performance liquid chromatography of an enzymatic peptidoglycan hydrolysate and fast atom bombardment mass spectrometry and fast atom bombardment collision-activated dissociation tandem mass spectrometry. Sixteen major components of the peptidoglycan were purified, and assignment of complete or partial chemical structures was achieved for nine and seven species, respectively. At this level of resolution, a previously unrecognized heterogeneity of monomeric (five new species; nine total) and dimeric species (five new species; five total) was detected. No species containing diaminopimelyl-diaminopimelic acid cross-links or lysyl-arginine substitutions were found. Previous estimates of total cross-linkage and average chain length were revised downward to 32% and 21 disaccharide residues, respectively. Detection of a chemically novel species, a disaccharide octapeptide monomer, in both the peptidoglycan hydrolysate and culture supernatant fluid, suggests that an N-acetyl-muramyl-L-alanine amidase acts on the intact peptidoglycan of Bordetella and participates in cell wall turnover. Five peptidoglycan turnover products were identified in the supernatant fluid of late logarithmic phase cultures, including the 1,6-anhydro monomeric species known as tracheal cytotoxin. Peptidoglycan turnover was detected at a low rate of approximately 10%/generation, a value sufficient to account for the generation of all tracheal cytotoxin found in culture supernatant fluids.
通过一种新的分析技术组合，即酶促肽聚糖水解产物的高效液相色谱法、快原子轰击质谱法和快原子轰击碰撞激活解离串联质谱法，确定了百日咳博德特氏菌肽聚糖的组成及其周转产物的性质。纯化了肽聚糖的16种主要成分，分别对9种和7种成分完成或部分完成了化学结构的鉴定。在这种分辨率水平下，检测到了单体（5个新种类；共9个）和二聚体种类（5个新种类；共5个）中以前未被识别的异质性。未发现含有二氨基庚二酸-二氨基庚二酸交联或赖氨酰-精氨酸取代的种类。先前对总交联度和平均链长的估计分别下调至32%和21个二糖残基。在肽聚糖水解产物和培养上清液中均检测到一种化学上新颖的种类，即二糖八肽单体，这表明N-乙酰胞壁酰-L-丙氨酸酰胺酶作用于百日咳博德特氏菌完整的肽聚糖并参与细胞壁周转。在对数生长期后期培养物的上清液中鉴定出5种肽聚糖周转产物，包括被称为气管细胞毒素的1,6-脱水单体种类。检测到肽聚糖周转的速率约为10%/代，该值足以解释培养上清液中发现的所有气管细胞毒素的产生。
