Wolters P J, Hildebrandt K M, Dickie J P, Anderson J S
Department of Biochemistry, College of Biological Sciences, University of Minnesota, St. Paul 55108.
J Bacteriol. 1990 Sep;172(9):5154-9. doi: 10.1128/jb.172.9.5154-5159.1990.
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.
通过温和酸处理从藤黄微球菌细胞壁中与肽聚糖的磷酸二酯键释放出来的磷壁醛酸,在硼酸盐存在下于聚丙烯酰胺凝胶上进行电泳时,会分解成一系列阶梯状条带。阶梯中的每条带与其相邻带相差一个二糖重复单元,即----4)-2-乙酰氨基-2-脱氧-β-D-甘露糖醛酸基-(1----6)-α-D-葡萄糖吡喃糖基-(1-。酸片段化的磷壁醛酸转化为苯胺衍生物后,通过制备规模的分子筛柱色谱进行分离,产生一系列洗脱峰。该系列中最小成员的快原子轰击质谱测定了其分子量,并确定其为磷壁醛酸一个二糖重复单元的苯胺衍生物。同一系列的同源级分用于对含有更广泛聚合物长度分布的样品通过聚丙烯酰胺凝胶电泳获得的条带阶梯进行索引。接近天然的磷壁醛酸由分子大小范围从20到55个二糖单元的聚合物组成。最丰富的种类是那些具有25到40个重复单元的聚合物。用用于从肽聚糖中释放磷壁醛酸的酸性条件对其进行长时间处理会导致磷壁醛酸逐渐片段化。