蛋白质精准修饰客体分子的方法  

Methodologies for the Precise Modification of Guest Molecules by Proteins

作　　者：戴震 刘育[1] 刘涛 DAI Zhen;LIU Yu;LIU Tao(College of Chemistry,State Key Laboratory of Element of Organic Chemistry,Nankai University,Tianjin 300071,China;State Key Laboratory of Natural and Biomimetic Drugs,School of Pharmaceutical Sciences,School of Pharmaceutical Sciences,Peking University,Beijing 100191,China;Chemical Biology Center,School of Pharmaceutical Sciences,Peking University,Beijing 100191,China;Department of Molecular and Cellular Pharmacology,School of Pharmaceutical Sciences,Peking University,Beijing 100191,China)

机构地区：[1]南开大学化学学院,元素有机化学国家重点实验室,天津300071 [2]北京大学药学院,天然药物及仿生药物国家重点实验室,北京100191 [3]北京大学药学院化学生物学交叉中心,北京100191 [4]北京大学药学院药学院分子与细胞药理学系,北京100191

出　　处：《高等学校化学学报》2024年第6期69-84,共16页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:22325701,U22A20332,92156025,92253301);国家重点研发计划项目(批准号:2022YFA0912400,2021YFA0909900);北京市自然科学基金(批准号:JQ20034)的资助。

摘　　要：客体分子在蛋白质上的精准修饰为超分子调控蛋白质活性提供了重要技术基础,尤其是带正电荷的芳香官能团客体,可以与葫芦脲高效地结合.然而,该类分子很难通过非天然氨基酸定点修饰技术精准修饰到蛋白质上,而生物正交反应的连接子通常又较大从而影响主客体分子识别.为此,本文通过非天然氨基酸定点修饰技术先将带碘或炔烃官能团的非天然氨基酸定点修饰到目的蛋白质上,再通过钯催化的Suzuki和Sonogashira偶联反应进一步修饰上带有正电荷的吡啶分子,以期与葫芦脲分子高效识别结合,通过对反应条件的筛选插入了带有炔烃官能团的非天然氨基酸,并通过Sonogashira偶联反应在蛋白质表面定点偶联带正电荷的吡啶分子,为主客体化学调控蛋白质奠定了重要的技术基础.Precise modification of guest molecules on proteins is a crucial technological foundation for supramolecular modulation of protein activity.This is especially true for aromatic functional group guests containing positive charges,which can be efficiently bound to cucurbituril.However,precise modification of these molecules on proteins using non-canonical amino acid site specific modification technology can be challenging.The linkers of bioorthogonal reactions are often large,which can affect the recognition of the host and guest molecules.To address this issue,the target proteins were modified by adding iodine-or alkynyl-functional amino acids using the non-canonical amino acid site specific modification technology.The pyridine molecules with a positive charge were efficiently bound to cucurbi⁃turil molecules using the palladium-catalyzed Suzuki and Sonogashira coupling reaction.Subsequently,a non-natural amino acid with an alkynyl functional group was successfully incorporated into the target proteins using a series of reaction conditions.Furthermore,a Sonogashira coupling reaction was utilized to attach a positively charged pyridine molecule to the protein surface.These accomplishments establish a crucial technological basis for the chemical regulation of proteins through host-guest interactions.

关 键 词：精准修饰 非天然氨基酸 主客体 蛋白质 

分 类 号：O621.3[理学—有机化学] O641.3[理学—化学]