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胱天蛋白酶3

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胱天蛋白酶3
已知的結構
PDB直系同源搜尋: PDBe RCSB
識別號
別名CASP3;, CPP32, CPP32B, SCA-1, caspase 3
外部IDOMIM600636 MGI107739 HomoloGene37912 GeneCardsCASP3
為以下藥物的標靶
emricasan[1]
RNA表達模式
查閱更多表達數據
直系同源
物種人類小鼠
Entrez
Ensembl
UniProt
mRNA​序列

​NM_004346
​NM_032991

NM_009810
​NM_001284409

蛋白序列

NP_001271338
​NP_033940

基因位置​(UCSC)無數據Chr 8: 47.07 – 47.09 Mb
PubMed​查找[3][4]
維基數據
檢視/編輯人類檢視/編輯小鼠

胱天蛋白酶3英語:Caspase 3)是一種在人類中由CASP3基因編碼的。該酶能與胱天蛋白酶8胱天蛋白酶9產生相互作用。許多可獲得完整基因組數據的哺乳動物都已鑑定出CASP3直系同源物[5]鳥類蜥蜴滑體動物真骨類中也存在獨特的直系同源物。

胱天蛋白酶3是胱天蛋白酶(Caspase)家族的成員。[6]胱天蛋白酶的連續活化在細胞凋亡的執行階段發揮着核心作用。胱天蛋白酶以無活性的酶原形式存在,在保守的天冬氨酸殘基處經歷蛋白水解加工,產生一大一小兩個亞基,然後二聚化形成活性酶。該蛋白酶可裂解並活化胱天蛋白酶67,其本身則由胱天蛋白酶8、9和10加工和活化。該蛋白酶是參與裂解前類澱粉蛋白質的主要胱天蛋白酶,而前類澱粉蛋白質與阿茲海默症中的神經元死亡有關。[7]該基因的選擇性剪接會產生編碼相同蛋白質的兩個轉錄變體。[8]

TNF-R1信號通路。灰色虛線代表多個步驟
導致胱天蛋白酶3活化的途徑[9]

胱天蛋白酶3具有許多目前已知的胱天蛋白酶共有的典型特徵。例如,其活性位點包含半胱氨酸殘基(Cys-163)和組氨酸殘基(His-121),當它位於特定的4個氨基酸序列中時,它能夠穩定蛋白質序列肽鍵裂解到天冬氨酸羧基末端側。[10][11]這種特異性使得胱天蛋白酶具有有極高的選擇性,對天冬氨酸的偏好是穀氨酸的2萬倍。[12]胱天蛋白酶在細胞中的一個關鍵特徵是它們以未活化的前酶形式存在,稱為胱天蛋白酶原,直到生化變化引起它們的活化為止。每個胱天蛋白酶原都有一個約20kDa的N端大亞基,後面跟着一個約10kDa的小亞基,分別稱為p20和p10。[13]

基質特異性

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正常情況下,胱天蛋白酶識別其基質上的四肽序列並水解天冬氨酸殘基後的肽鍵。胱天蛋白酶3和7通過識別四肽基序Asp-x-x-Asp共享類似的基質特異性。[14]C端天冬氨酸是絕對必需的,而其他三個位置的變化是可以容忍的。[15]胱天蛋白酶基質特異性已廣泛應用於基於胱天蛋白酶的抑制劑和藥物設計。[16]

結構

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subunits alt text
胱天蛋白酶3的p12(粉色)和 p17(淺藍色)亞基,其β摺疊結構分別為紅色和藍色;在Pymol中從1rhm.pdb生成的圖像

胱天蛋白酶3(也稱為CPP32、Yama或apopain)[17][18][19]是由一個32kDa的酶原形成,該酶原被切割成17kDa和12kDa亞基。當胱天蛋白酶原在特定殘基處裂解時,活性異四聚體就能通過疏水相互作用形成,導致來自p17的四個反平行β摺疊和來自p12的兩個反平行β摺疊結合在一起形成異二聚體,該異二聚體又與另一個異二聚體相互作用形成完整的由α螺旋包圍的12鏈β摺疊結構,這是胱天蛋白酶特有的。[13][20]當異二聚體頭尾相連時,分子兩端各有一個由兩個參與亞基的殘基形成的活性位點,儘管必要的Cys-163和His-121殘基位於p17(較大的)亞基上。[20]

機制

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active site alt text
胱天蛋白酶3活性位點的Cys-285(黃色)和His-237(綠色和深藍色),p12亞基為粉色,p17亞基為淺藍色;在Pymol中從1rhm.pdb生成的圖像

胱天蛋白酶3的催化位點涉及Cys-163的硫醇基團和His-121的咪唑環。His-121穩定關鍵天冬氨酸殘基的羰基,而Cys-163則攻擊最終裂解肽鍵。Cys-163和Gly-238還可以通過氫鍵穩定基質-酶複合物的四面體過渡態[20]在體外,已發現胱天蛋白酶3更喜歡肽序列 DEVDG(Asp-Glu-Val-Asp-Gly),其切割發生在第二個天冬氨酸殘基的羧基側(D和G之間)。[12][20][21]胱天蛋白酶3在較寬的pH範圍內具有活性,該範圍比許多其他執行型胱天蛋白酶稍高(鹼性更強)。這一廣泛的範圍表明胱天蛋白酶3在正常和凋亡細胞條件下都可以完全活躍。[22]

活化

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胱天蛋白酶3在凋亡細胞中通過外在(死亡配體)和內在(線粒體)途徑被活化。[13][23]胱天蛋白酶3的酶原特徵是必要的,因為如果不受調節,胱天蛋白酶的活性會不加區別地殺死細胞。[24]作為執行型胱天蛋白酶,胱天蛋白酶3酶原實際上沒有活性,直到凋亡信號事件發生後被啟動型胱天蛋白酶切割。[25]此類信號事件之一是將顆粒酶B引入殺傷性T細胞針對凋亡的細胞中,該顆粒酶B可以活化啟動型胱天蛋白酶。[26][27]這種外在活化隨後觸發細胞凋亡途徑的標誌性胱天蛋白酶級聯特徵,其中胱天蛋白酶3發揮主導作用。[11]在內在活化過程中,來自線粒體的細胞色素c與胱天蛋白酶9、凋亡活化因子1(Apaf-1)和ATP結合作用來處理胱天蛋白酶3酶原。[21][27][28]這些分子足以在體外活化胱天蛋白酶3,但體內還需要其他調節蛋白。[28]山竹Garcinia mangostana)提取物已被證明可以抑制β澱粉樣蛋白處理的人類神經元細胞中胱天蛋白酶3的活化。[29]

抑制

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抑制胱天蛋白酶的一種方法是通過IAP(凋亡抑制劑)蛋白家族,其中包括c-IAP1、c-IAP2、XIAP和ML-IAP。[20]XIAP結合併抑制啟動型胱天蛋白酶9,後者直接參與執行型胱天蛋白酶3的活化。[28]然而,在胱天蛋白酶級聯過程中,胱天蛋白酶3通過在特定位點切割胱天蛋白酶9來抑制XIAP的活性,從而阻止XIAP結合來抑制胱天蛋白酶9的活性。[30]

相互作用

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胱天蛋白酶3已被證明可以與以下物質相互作用

生物學功能

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人們發現胱天蛋白酶3對於正常的大腦發育是必需的,它在細胞凋亡中也發揮着典型作用,負責染色質濃縮和DNA碎裂。[21]血液中胱天蛋白酶3片段(p17)水平升高是近期心肌梗塞的徵兆。[52]現在的研究表明,胱天蛋白酶3可能在胚胎和造血幹細胞分化中發揮作用。[53]

參見

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參考文獻

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