Purification and characterization of a cysteine-rich 11.5-kDa antibacterial protein from the granular haemocytes of the shore crab, Carcinus maenas
Purification and characterization of a cysteine-rich 11.5-kDa antibacterial protein from the granular haemocytes of the shore crab, Carcinus maenas
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DOI：
10.1046/j.1432-1327.1999.00607.x
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发表时间：
1999-09-01
期刊：
影响因子：
--
通讯作者：
Smith, VJ
中科院分区：
文献类型：
--
作者：
Relf, JM;Chisholm, JRS;Smith, VJ
采用离子交换色谱法和反相高效液相色谱法（RP -HPLC）研究了一种约11 kDa的抗菌蛋白的存在，并对该蛋白进行了分离、表征和部分氨基酸序列分析。质谱分析发现其分子质量为11 534 Da，具有阳离子疏水性，仅对海洋革兰氏阳性菌有活性。此外，其活性在加热到100℃后保持稳定，并在低至10 μ g.mL（-1）的浓度下保持不变。它有一个不寻常的氨基酸序列，不像文献中描述的任何已知的抗菌肽，但具有一致的二硫结构域特征，表明它可能是四二硫核心蛋白的成员。获得了部分cDNA序列数据。
Extracts of the granular haemocytes of Carcinus maenas were subjected to ion-exchange chromatography and reverse-phase (RP)-HPLC to investigate the presence of an antibacterial protein of approximate to 11 kDa, This protein was isolated, characterized and subjected to partial amino acid sequence analysis. It was found by mass spectrometry to have a molecular mass of 11 534 Da, to be cationic and hydrophobic and active only against marine Gram-positive bacteria. In addition its activity is stable after heating to 100 degrees C and is retained at concentrations as low as 10 mu g.mL(-1). It has an unusual amino acid sequence, unlike any known antibacterial peptide described in the literature but bears a consensus disulphide domain signature, indicating that it might be a member of the four-disulphide core proteins. Partial cDNA sequence data has been obtained.