蓖麻毒素和相思豆毒素核糖体失活蛋白结构中的过渡态类似物。
Transition state analogues in structures of ricin and saporin ribosome-inactivating proteins.
机构信息
Department of Biochemistry, Albert Einstein College of Medicine, Yeshiva University, Bronx, NY 10461, USA.
出版信息
Proc Natl Acad Sci U S A. 2009 Dec 1;106(48):20276-81. doi: 10.1073/pnas.0911606106. Epub 2009 Nov 17.
Abstract
Ricin A-chain (RTA) and saporin-L1 (SAP) catalyze adenosine depurination of 28S rRNA to inhibit protein synthesis and cause cell death. We present the crystal structures of RTA and SAP in complex with transition state analogue inhibitors. These tight-binding inhibitors mimic the sarcin-ricin recognition loop of 28S rRNA and the dissociative ribocation transition state established for RTA catalysis. RTA and SAP share unique purine-binding geometry with quadruple pi-stacking interactions between adjacent adenine and guanine bases and 2 conserved tyrosines. An arginine at one end of the pi-stack provides cationic polarization and enhanced leaving group ability to the susceptible adenine. Common features of these ribosome-inactivating proteins include adenine leaving group activation, a remarkable lack of ribocation stabilization, and conserved glutamates as general bases for activation of the H(2)O nucleophile. Catalytic forces originate primarily from leaving group activation evident in both RTA and SAP in complex with transition state analogues.
摘要
蓖麻毒素 A 链（RTA）和丝氨酸蛋白酶 L1（SAP）催化腺苷脱嘌呤，抑制蛋白质合成并导致细胞死亡。我们展示了 RTA 和 SAP 与过渡态类似物抑制剂复合物的晶体结构。这些紧密结合的抑制剂模拟了 28S rRNA 的 sarcin-ricin 识别环和为 RTA 催化建立的解离核糖正离子过渡态。RTA 和 SAP 具有独特的嘌呤结合几何形状，相邻腺嘌呤和鸟嘌呤碱基之间存在四重 π-堆积相互作用，还有 2 个保守的酪氨酸。π-堆叠的一端的精氨酸提供阳离子极化和增强离去基团能力，使易受影响的腺嘌呤离去。这些核糖体失活蛋白的共同特征包括腺嘌呤离去基团的激活、核糖正离子的稳定作用显著缺乏以及保守的谷氨酸作为激活 H（2）O 亲核试剂的通用碱基。催化力主要源自离去基团的激活，这在 RTA 和 SAP 与过渡态类似物复合物中都很明显。
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