Harris Paul W R, Yang Sung-Hyun, Molina Antonio, López Gemma, Middleditch Martin, Brimble Margaret A
School of Chemical Sciences, The University of Auckland, 23 Symonds St., Auckland 1142 (New Zealand), Fax: (+64) 93737422; Maurice Wilkins Centre for Molecular Biodiscovery, School of Biological Sciences, The University of Auckland, Auckland 1142 (New Zealand).
Chemistry. 2014 Apr 22;20(17):5102-10. doi: 10.1002/chem.201303207. Epub 2014 Mar 18.
Antimicrobial peptides and proteins represent an important class of plant defensive compounds against pathogens and provide a rich source of lead compounds in the field of drug discovery. We describe the effective preparation of the cysteine-rich snakin-1 and -2 antimicrobial peptides by using a combination of solid-phase synthesis and native chemical ligation. A subsequent cysteine/cystine mediated oxidative folding to form the six internal disulfide bonds concurrently gave the folded proteins in 40-50 % yield. By comparative evaluation of mass spectrometry, HPLC, biological data and trypsin digest mapping of folded synthetic snakin-2 compared to natural snakin-2, we demonstrated that synthetic snakin-2 possesses full antifungal activity and displayed similar chromatographic behaviour to natural snakin-2. Trypsin digest analysis allowed tentative assignment of three of the purported six disulfide bonds.
抗菌肽和蛋白质是植物抵御病原体的一类重要防御化合物，也是药物发现领域中先导化合物的丰富来源。我们描述了通过固相合成和天然化学连接相结合的方法有效制备富含半胱氨酸的蛇形抗菌肽-1和-2。随后通过半胱氨酸/胱氨酸介导的氧化折叠同时形成六个内部二硫键，以40%-50%的产率得到折叠后的蛋白质。通过对折叠后的合成蛇形抗菌肽-2与天然蛇形抗菌肽-2进行质谱、高效液相色谱、生物学数据和胰蛋白酶消化图谱的比较评估，我们证明合成蛇形抗菌肽-2具有完全的抗真菌活性，并且在色谱行为上与天然蛇形抗菌肽-2相似。胰蛋白酶消化分析初步确定了六个假定二硫键中的三个。
