高专一性胰蛋白酶突变体的分子设计

本文对于改变胰蛋白酶专一性的方法进行了研究,通过对胰蛋白酶静电性质的计算找到了一种改变表面碱性氨基酸来达到使酶选择性地与精氨酸底物反应的途径,并给出了推荐的突变体表列。     

Molecular Design of Trypsin Towards High Substrate Selectivity

Molecular design of trypsin mutants towards higher substrate specificity for arginine or ly-sine type substrates was studied. The difference in side chain pKa of arginine and lysine was utilized in redesigning trypsin. If the enzyme could react effectively at a pH higher than lysine's pKa but lower than that of arginine, it would react more selectively with arginine-type substrates, since in that pH range, the side chain of arginie remain protonated, while that of lysine is deprotonated. For trypsin. the change of histidine (57)'s pKa reflects the shift in reaction optimum pH. Electrostatic calculations showed that when surface positive residues were mutated into neutral or negative ones, the pKa of histidine(57) would be raised and those surface charges within a cone of 70 degree around histidine(57) have strong influence on its pKa. Several sites were suggested in rat trypsin which might serve as potential mutation locations to make trypsin active at a higher pH, thus more selective towards arginine t