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我國科學家為活細胞蛋白質安裝“調控開關”

文章來源:光明日報 晉浩天   發布時間:2019-05-10  【字號:     】  

  北京大學化學與分子工程學院、合成與功能生物分子中心陳鵬課題組與王初課題組,8日在《自然》雜志在線發表題為《基于鄰近脫籠技術的活體內蛋白質瞬時激活》的研究論文,報道了兩個課題組合作發展的一種在活體環境下瞬時激活蛋白質的化學生物學新技術。

  在活細胞等生理環境下開展蛋白質功能的原位研究具有重要的科學意義。陳鵬課題組長期致力于發展蛋白質的原位激活技術,希望為活細胞內的每一個蛋白質安裝“調控開關”。最近,他們與王初課題組展開合作,提出并發展了一種蛋白質“鄰近脫籠”策略,將可遺傳編碼的非天然氨基酸脫籠技術與計算機輔助設計篩選技術相結合,在一系列不同種類的蛋白質上實現了高時間分辨的原位激活,為在活細胞及活體動物內研究蛋白質的動態調控機制提供了一種普適性技術。利用這一被命名為CAGE-prox的新方法,他們建立并驗證了“激酶正交激活和信號轉導調控”“時間分辨的蛋白質組學分析”“基于毒素蛋白的抗腫瘤蛋白前藥”等一系列原創應用,展示了這一化學生物學新技術在開展蛋白質動態功能研究與調控中的優勢和特色。

  陳鵬課題組先前提出的“化學脫籠”策略,可通過對蛋白質關鍵殘基的化學保護和脫保護反應,實現對其活性的“關-開”調控。利用這一技術,他們先后在賴氨酸、酪氨酸等天然氨基酸側鏈上實現了生物正交斷鍵反應和化學脫籠,開展了激酶等蛋白質家族的特異激活和機制研究。由于細胞內蛋白質的種類繁多、活性調控機制各異,目前該方法仍受可供脫籠的氨基酸種類限制,無法適用于所有蛋白質。

  為了解決這一問題,陳鵬課題組與王初課題組合作提出了“鄰近脫籠”的新策略,通過向蛋白質活性中心附近引入一種帶有光保護基團的非天然酪氨酸,可同樣實現對其活性的遠程干擾和抑制;隨后通過“光脫籠”反應將保護基團移除,使其活性重新恢復。由于在蛋白質活性口袋附近插入非天然酪氨酸后能夠通過影響蛋白穩定性、底物結合等多種因素調控蛋白質活性,該策略將原有的“活性位點脫籠”轉變為“活性口袋脫籠”,極大地擴展了該方法的適用范圍。

  中科院院士、北京大學醫學部教授張禮和表示,陳鵬課題組此前率先利用基于生物正交斷鍵反應的化學脫籠策略,結合遺傳密碼子拓展技術,實現了在蛋白質催化位點的生物正交脫籠,進而原位激活其功能。在最新的工作中,陳鵬課題組與北大王初課題組合作,進一步提出并發展了“鄰近脫籠”的新策略,利用計算機輔助設計與篩選,將蛋白質脫籠技術的適用范圍從催化位點拓展至整個活性口袋,獲得了具有廣泛適用性的蛋白質激活新方法。該工作是化學生物學領域的一項重要進展,為在活細胞及活體動物內開展蛋白質動態功能研究提供了關鍵技術,也為生物正交反應開拓了新的前沿方向。

  據悉,北京大學前沿交叉學科研究院博士畢業生王杰、化學與分子工程學院博士后劉源、研究生劉衍軍為該論文共同第一作者,陳鵬教授和王初研究員為本文的共同通訊作者。該項工作得到了國家自然科學基金委、科技部、教育部、北京分子科學國家研究中心和北大—清華生命科學聯合中心的資助。




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