Departamento de Biologia Molecular, Universidade Federal da Paraíba, João Pessoa, Brazil.
Appl Biochem Biotechnol. 2011 Jul;164(6):741-54. doi: 10.1007/s12010-011-9170-x. Epub 2011 Feb 22.
The lectin from seeds of Dioclea virgata (DvirL) was purified in a single step affinity chromatography, sequenced by tandem mass spectrometry and submitted to crystallization and biological experiments. DvirL has a molecular mass of 25,412 ± 2 Da and the chains β and γ has 12,817 Da ± 2 and 12,612 Da ± 2, respectively. Primary sequence determination was assigned by tandem mass spectrometry and revealed a protein with 237 amino acids and 87% of identify with ConA. The protein crystals were obtained native and complexed with X-Man using vapor-diffusion method at a constant temperature of 293 K. A complete X-ray dataset was collected at 1.8 Å resolution. DvirL crystals were found to be orthorhombic, belonging to the space group I222, with a unit cell parameters a = 647.5 Å, b = 86.6 Å, c = 90.2 Å. Molecular replacement search found a solution with a correlation coefficient of 77.1% and an R(factor) of 44.6%. The present study also demonstrated that D. virgata lectin presents edematogenic and antinociceptive activities in rodents electing this protein as a candidate to structure/function analysis.
从 Dioclea virgata 种子中纯化的凝集素（DvirL）通过一步亲和层析进行纯化，通过串联质谱法进行测序，并进行结晶和生物学实验。DvirL 的分子量为 25412±2 Da，β 和 γ 链分别为 12817 Da±2 和 12612 Da±2。通过串联质谱法确定了一级序列测定，揭示了一种具有 237 个氨基酸的蛋白质，与 ConA 的同一性为 87%。使用蒸汽扩散法在 293 K 的恒定温度下获得了天然和与 X-Man 复合的蛋白质晶体。在 1.8 Å 的分辨率下收集了完整的 X 射线数据集。DvirL 晶体呈正交晶系，属于空间群 I222，具有单位晶胞参数 a=647.5 Å、b=86.6 Å、c=90.2 Å。分子置换搜索发现了一个相关系数为 77.1%、R(factor)为 44.6%的解决方案。本研究还表明，D. virgata 凝集素在啮齿动物中具有水肿和镇痛活性，选择这种蛋白质作为结构/功能分析的候选物。
