Agerberth B, Lee J Y, Bergman T, Carlquist M, Boman H G, Mutt V, Jörnvall H
Department of Biochemistry II, Karolinska Institutet, Stockholm, Sweden.
Eur J Biochem. 1991 Dec 18;202(3):849-54. doi: 10.1111/j.1432-1033.1991.tb16442.x.
We recently isolated from pig intestine and characterized a 31-residue antibacterial peptide named cecropin-P1 with activity against Escherichia coli and several other Gram-negative bacteria. The isolation involved a number of batch-wise steps followed by several chromatography steps. The continued investigation of these antibacterial peptides has now yielded another antibacterial peptide with high activity against both E. coli and Bacillus megaterium. Amino acid analysis showed a very high content of proline (49 mol%) and arginine (26 mol%), an intermediate level of phenylalanine and low levels of leucine, tyrosine, isoleucine, and glycine. The primary structure was determined by a combination of Edman degradation, plasma desorption mass spectrometry and C-terminal sequence analysis by carboxypeptidase Y degradation using capillary zone electrophoresis for detection of liberated residues. The calculated molecular mass was 4719.7 Da, which is in excellent agreement with 4719 Da obtained by plasma desorption mass spectrometry. The peptide was named PR-39 (proline-arginine-rich with a size of 39 residues). The lethal concentration of the peptide was determined against six Gram-negative and four Gram-positive strains of bacteria.
我们最近从猪肠道中分离出一种名为天蚕素 - P1的31个氨基酸残基的抗菌肽，并对其进行了表征，该抗菌肽对大肠杆菌和其他几种革兰氏阴性菌具有活性。分离过程包括多个分批步骤，随后是几个色谱步骤。对这些抗菌肽的持续研究现已产生了另一种对大肠杆菌和巨大芽孢杆菌均具有高活性的抗菌肽。氨基酸分析表明，脯氨酸含量非常高（49摩尔%），精氨酸含量较高（26摩尔%），苯丙氨酸含量处于中等水平，亮氨酸、酪氨酸、异亮氨酸和甘氨酸含量较低。通过埃德曼降解、等离子体解吸质谱以及使用毛细管区带电泳检测释放残基的羧肽酶Y降解进行C端序列分析相结合的方法确定了其一级结构。计算得到的分子量为4719.7 Da，与通过等离子体解吸质谱获得的4719 Da非常吻合。该肽被命名为PR - 39（富含脯氨酸 - 精氨酸，大小为39个残基）。测定了该肽对六种革兰氏阴性菌和四种革兰氏阳性菌菌株的致死浓度。
