Costa M H B, Quintilio W, Sant'Anna O A, Faljoni-Alário A, de Araujo P S
Laboratório de Microesferas e Lipossomos, Centro de Biotecnologia, Instituto Butantan, São Paulo, SP, Brasil.
Braz J Med Biol Res. 2002 Jun;35(6):727-30. doi: 10.1590/s0100-879x2002000600014.
The recombinant heat shock protein (18 kDa-hsp) from Mycobacterium leprae was studied as a T-epitope model for vaccine development. We present a structural analysis of the stability of recombinant 18 kDa-hsp during different processing steps. Circular dichroism and ELISA were used to monitor protein structure after thermal stress, lyophilization and chemical modification. We observed that the 18 kDa-hsp is extremely resistant to a wide range of temperatures (60% of activity is retained at 80 degrees C for 20 min). N-Acylation increased its ordered structure by 4% and decreased its beta-T1 structure by 2%. ELISA demonstrated that the native conformation of the 18 kDa-hsp was preserved after hydrophobic modification by acylation. The recombinant 18 kDa-hsp resists to a wide range of temperatures and chemical modifications without loss of its main characteristic, which is to be a source of T epitopes. This resistance is probably directly related to its lack of organization at the level of tertiary and secondary structures.
对来自麻风分枝杆菌的重组热休克蛋白（18 kDa - hsp）作为疫苗开发的T表位模型进行了研究。我们对重组18 kDa - hsp在不同加工步骤中的稳定性进行了结构分析。利用圆二色性和酶联免疫吸附测定法监测热应激、冻干和化学修饰后的蛋白质结构。我们观察到，18 kDa - hsp对广泛的温度具有极强的抗性（在80℃下20分钟仍保留60%的活性）。N - 酰化作用使其有序结构增加了4%，β - T1结构减少了2%。酶联免疫吸附测定法表明，经酰化进行疏水修饰后，18 kDa - hsp的天然构象得以保留。重组18 kDa - hsp能抵抗广泛的温度和化学修饰，且不丧失其作为T表位来源的主要特性。这种抗性可能与其在三级和二级结构水平上缺乏有序组织直接相关。
