Semisynthesis of Aminomethyl-Insulin: An Atom-Economic Strategy to Increase the Affinity and Selectivity of a Protein for Recognition by a Synthetic Receptor.
Semisynthesis of Aminomethyl-Insulin: An Atom-Economic Strategy to Increase the Affinity and Selectivity of a Protein for Recognition by a Synthetic Receptor.
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DOI：
10.1021/acs.bioconjchem.2c00501
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发表时间：
2023-01-18
影响因子：
4.7
通讯作者：
Urbach AR
中科院分区：
文献类型：
--
作者：
Anderson HR;Reeves WL;Bockus AT;Suating P;Grice AG;Gallagher M;Urbach AR
通过非抗体方法识别胰岛素分子的进展将有助于开发用于糖尿病管理的胰岛素连续检测方法。在此，我们报告了一种新的胰岛素衍生物，它与合成受体葫芦[7]脲（Q7）在一个单一的位点结合，并具有高纳摩尔亲和力。通过四步蛋白质半合成方法制备胰岛素衍生物，以在PheB 1位置的侧链上呈现4-氨甲基。通过等温滴定量热法测定，所得氨甲基胰岛素与Q7结合，在中性磷酸盐缓冲液中的平衡解离常数值为99 nM。与天然胰岛素相比，亲和力提高了6.8倍，这是通过原子经济性修饰（− CH 2NH 2）获得的。据我们所知，这是合成受体对胰岛素衍生物的最高报告结合亲和力。这种用于工程化蛋白质亲和标签的策略诱导蛋白质结构的最小变化，同时增加对合成受体的亲和力和选择性。
Advancements in the molecular recognition of insulin by nonantibody-based means would facilitate the development of methodology for the continuous detection of insulin for the management of diabetes mellitus. Herein, we report a novel insulin derivative that binds to the synthetic receptor cucurbit[7]uril (Q7) at a single site and with high nanomolar affinity. The insulin derivative was prepared by a four-step protein semisynthetic method to present a 4-aminomethyl group on the side chain of the PheB1 position. The resulting aminomethyl insulin binds to Q7 with an equilibrium dissociation constant value of 99 nM in neutral phosphate buffer, as determined by isothermal titration calorimetry. This 6.8-fold enhancement in affinity versus native insulin was gained by an atom-economical modification (−CH2NH2). To the best of our knowledge, this is the highest reported binding affinity for an insulin derivative by a synthetic receptor. This strategy for engineering protein affinity tags induces minimal change to the protein structure while increasing affinity and selectivity for a synthetic receptor.
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影响因子：
56.9
作者：
DAWSON, PE;MUIR, TW;KENT, SBH
通讯作者：
KENT, SBH
影响因子：
1.8
作者：
Ali, Asghar;van den Berg, Richard J. B. H. N.;Codee, Jeroen D. C.
通讯作者：
Codee, Jeroen D. C.
影响因子：
4.9
作者：
Li, Yijia;Zhao, Linlu;Liu, Junqiu
通讯作者：
Liu, Junqiu
影响因子：
15
作者：
Avital-Shmilovici, Michal;Mandal, Kalyaneswar;Gates, Zachary P.;Phillips, Nelson B.;Weiss, Michael A.;Kent, Stephen B. H.
通讯作者：
Kent, Stephen B. H.
影响因子：
15
作者：
Logsdon, Leigh A.;Schardon, Christopher L.;Urbach, Adam R.
通讯作者：
Urbach, Adam R.