Lipkin Aleksey, Anisimova Veronika, Nikonorova Aleksandra, Babakov Aleksey, Krause Eberhardt, Bienert Mikhael, Grishin Eugene, Egorov Tsezi
Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow.
Phytochemistry. 2005 Oct;66(20):2426-31. doi: 10.1016/j.phytochem.2005.07.015. Epub 2005 Aug 29.
A 30-residue antimicrobial peptide Ar-AMP was isolated from the seeds of amaranth Amaranthus retroflexus L. essentially by a single step procedure using reversed-phase HPLC, and its in vitro biological activities were studied. The complete amino acid sequence of Ar-AMP was determined by Edman degradation in combination with mass spectrometric methods. In addition, the cDNA encoding Ar-AMP was obtained and sequenced. The cDNA encodes a precursor protein consisting of the N-terminal putative signal sequence of 25 amino acids, a mature peptide of 30 amino acids and a 34-residue long C-terminal region cleaved during post-translational processing. According to sequence similarity the Ar-AMP belongs to the hevein-like family of antimicrobial peptides with six cysteine residues. In spite of the fact that seeds were collected in 1967 and lost their germination capacity, Ar-AMP retained its biological activities. It effectively inhibited the growth of different fungi tested: Fusarium culmorium (Smith) Sacc., Helminthosporium sativum Pammel., King et Bakke, Alternaria consortiale Fr., and Botrytis cinerea Pers., caused morphological changes in Rhizoctonia solani Kühn at micromolar concentrations and protected barley seedlings from H. sativum infection.
一种由30个氨基酸残基组成的抗菌肽Ar-AMP,主要通过反相高效液相色谱法单步从反枝苋(Amaranthus retroflexus L.)种子中分离得到,并对其体外生物活性进行了研究。Ar-AMP的完整氨基酸序列通过埃德曼降解法结合质谱法确定。此外,获得了编码Ar-AMP的cDNA并进行了测序。该cDNA编码一种前体蛋白,其由25个氨基酸的N端假定信号序列、30个氨基酸的成熟肽和一个在翻译后加工过程中被切割的34个氨基酸残基的C端区域组成。根据序列相似性,Ar-AMP属于具有六个半胱氨酸残基的类橡胶素抗菌肽家族。尽管种子于1967年采集且已失去发芽能力,但Ar-AMP仍保留其生物活性。它能有效抑制所测试的不同真菌的生长:禾谷镰刀菌(Fusarium culmorium (Smith) Sacc.)、小麦根腐病菌(Helminthosporium sativum Pammel.、King及Bakke)、链格孢菌(Alternaria consortiale Fr.)和灰葡萄孢菌(Botrytis cinerea Pers.),在微摩尔浓度下引起立枯丝核菌(Rhizoctonia solani Kühn)的形态变化,并保护大麦幼苗免受小麦根腐病菌的感染。