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Abstract：
Teichoic acids (TA) are anionic polymers comprised of polyol phosphate repeat units that are abundant in the cell wall of Gram-positive bacteria. Both wall teichoic acid (WTA) and lipoteichoic acid (LTA) play important roles in regulating cell wall remodeling as well as conferring antibiotic resistance. To analyze TA, we describe a polyacrylamide gel electrophoresis (PAGE) method for both WTA and LTA. To extract crude WTA, the peptidoglycan sacculus is first isolated and WTA is then liberated by hydrolysis. LTA is extracted by 1-butanol and pre-treated with lipase to prevent aggregation and improve single-band resolution by PAGE. Crude extracts of both TAs are then subjected to PAGE followed by Alcian blue and silver staining. These protocols are easily adoptable by laboratories interested in rapidly analyzing TAs and can be used determine the relative abundance, relative polymer length and whether TAs are glycosylated. More detailed TA structural and compositional information can be obtained using the described purification protocols by nuclear magnetic resonance (NMR) and electrospray ionization mass spectrometry (ESI-MS) analysis.
摘要：
磷壁酸(TA)是由在革兰氏阳性细菌的细胞壁中丰富的多元醇磷酸酯重复单元组成的阴离子聚合物。壁磷壁酸（WTA）和脂磷壁酸（LTA）在调节细胞壁重塑以及赋予抗生素抗性中起重要作用。要分析TA，我们描述了WTA和LTA的聚丙烯酰胺凝胶电泳(PAGE)方法。为了提取粗WTA，首先分离肽聚糖囊,然后通过水解释放WTA。LTA通过1-丁醇提取并用脂肪酶预处理以防止聚集并通过PAGE提高单带分辨率。然后对两种TA的粗提取物进行PAGE,随后进行阿尔辛蓝和银染色。这些协议很容易被有兴趣快速分析TA的实验室采用，可以用来确定相对丰度。相对聚合物长度和TA是否糖基化。可以使用所述纯化方案通过核磁共振(NMR)和电喷雾电离质谱(ESI-MS)分析获得更详细的TA结构和组成信息。
