PENTZ E I, KOT E, FERRETTI J J
J Bacteriol. 1964 Aug;88(2):497-508. doi: 10.1128/jb.88.2.497-508.1964.
Pentz, E. Irene (Northwestern University School of Medicine, Chicago, Ill.), Eva Kot, and J. J. Ferretti. Some characteristics of streptococcal nicotinamide adenine dinucleotidase and streptolysin O. J. Bacteriol. 88:497-508. 1964.-Nicotinamide adenine dinucleotidase produced by Streptococcus pyogenes strain C203U, grown in Todd-Hewitt broth (Difco), was recovered from diethylaminoethyl-cellulose columns free from streptolysin O. It contained a trace of deoxyribonuclease. The elution peak of nicotinamide adenine dinucleotidase was fairly symmetrical; when it was treated with dinitrofluorobenzene (DNFB) and chromatogrammed on paper in three different solvent systems, only one yellow spot was obtained. Examination of the enzyme in an ultracentrifuge presented no moving boundary. The pore size of two different widths of Visking casing was estimated by dialyzing proteins or peptides of known molecular weight and testing the dialysate for their appearance. The casing which allowed a molecule of 2,500 molecular weight to pass, but not one of 3,000 molecular weight, retained the nicotinamide adenine dinucleotidase. A fragment having no nicotinamide adenine dinucleotidase activity did pass through this casing, however, but was retained by the second casing which retained molecules weighing 2,500. Amino acid analyses of three different nicotinamide adenine dinucleotidase preparations were remarkably similar, and were characterized by a high content of proline and glycine and a very low content of sulfur-containing amino acids. Examination by treatment with DNFB followed by hydrolysis for the presence of N-terminal amino acids ruled out all amino acids, with the possible exception of arginine. The combined evidence suggests that its molecular weight is between 2,500 and 20,000. Some data are presented to illustrate the irreversible instability of streptolysin O; amino acid analyses for three separate preparations of high activity indicate a very low content of sulfur-containing amino acids.
彭茨，艾琳·E.（伊利诺伊州芝加哥西北大学医学院），伊娃·科特，以及J.J.费雷蒂。化脓性链球菌烟酰胺腺嘌呤二核苷酸酶和链球菌溶血素O的一些特性。《细菌学杂志》88:497 - 508。1964年。——在托德 - 休伊特肉汤（迪夫科公司）中培养的化脓性链球菌C203U菌株产生的烟酰胺腺嘌呤二核苷酸酶，从不含链球菌溶血素O的二乙氨基乙基纤维素柱上回收。它含有微量的脱氧核糖核酸酶。烟酰胺腺嘌呤二核苷酸酶的洗脱峰相当对称；用二硝基氟苯（DNFB）处理并在三种不同溶剂系统中进行纸层析时，只得到一个黄色斑点。在超速离心机中对该酶进行检测未出现移动界面。通过透析已知分子量的蛋白质或肽并检测透析液中它们的出现情况，估计了两种不同宽度的维斯金肠衣的孔径。能使分子量为2500的分子通过但不能使分子量为3000的分子通过的肠衣，保留了烟酰胺腺嘌呤二核苷酸酶。然而，一个没有烟酰胺腺嘌呤二核苷酸酶活性的片段确实通过了这种肠衣，但被保留分子量为2500的分子的第二种肠衣所保留。对三种不同的烟酰胺腺嘌呤二核苷酸酶制剂进行的氨基酸分析非常相似，其特点是脯氨酸和甘氨酸含量高，含硫氨基酸含量极低。用DNFB处理后进行水解以检测N端氨基酸的存在，排除了所有氨基酸，可能除了精氨酸。综合证据表明其分子量在2500至20000之间。给出了一些数据来说明链球菌溶血素O的不可逆不稳定性；对三种高活性的单独制剂进行的氨基酸分析表明含硫氨基酸含量极低。
