Markham George D, Takusagawa Fusao, Dijulio Anthony M, Bock Charles W
Institute for Cancer Research, Fox Chase Cancer Center, 333 Cottman Avenue, Philadelphia, PA 19111, USA.
Arch Biochem Biophys. 2009 Dec;492(1-2):82-92. doi: 10.1016/j.abb.2009.08.010. Epub 2009 Aug 20.
Catalysis by S-adenosylmethionine synthetase has been investigated by quantum mechanical/molecular mechanical calculations, exploiting structures of the active crystalline enzyme. The transition state energy of +19.1 kcal/mol computed for a nucleophilic attack of the methionyl sulfur on carbon-5' of the nucleotide was indistinguishable from the experimental (solution) value when the QM residues were an uncharged histidine that hydrogen bonds to the leaving oxygen-5' and an aspartate that chelates a Mg2+ ion, and was similar (+18.8 kcal/mol) when the QM region also included the active site arginine and lysines. The computed energy difference between reactant and product was also consistent with their equimolar abundance in co-crystals. The calculated geometrical changes support catalysis of a S(N)2 reaction through hydrogen bonding of the liberated oxygen-5' to the histidine, charge neutralization by the two Mg2+ ions, and stabilization of the product sulfonium cation through a close, non-bonded, contact between the sulfur and the ribose oxygen-4'.
利用活性晶体酶的结构,通过量子力学/分子力学计算对S-腺苷甲硫氨酸合成酶的催化作用进行了研究。当量子力学(QM)残基为与离去的5'-氧形成氢键的不带电荷的组氨酸和螯合Mg2+离子的天冬氨酸时,针对甲硫氨酰硫对核苷酸5'-碳的亲核攻击计算出的+19.1千卡/摩尔的过渡态能量与实验(溶液)值无明显差异;当QM区域还包括活性位点精氨酸和赖氨酸时,该能量与之相似(+18.8千卡/摩尔)。计算出的反应物与产物之间的能量差也与它们在共晶体中的等摩尔丰度一致。计算出的几何变化支持通过将释放的5'-氧与组氨酸形成氢键、两个Mg2+离子进行电荷中和以及通过硫与核糖4'-氧之间紧密的非键接触来稳定产物锍阳离子,从而催化SN2反应。