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表觀遺傳修飾閱讀器”reader”蛋白結(jié)構(gòu)域靶點(diǎn)的發(fā)現(xiàn)

2021-05-17
瀏覽次數(shù): 2161
*注解:?將能識(shí)別特異性表觀修飾并與其結(jié)合的蛋白喻為“讀者或閱讀器(readers)”

眾所周知,小分子物質(zhì)很難抑制蛋白質(zhì)相互作用(Protein-Protein?Interactions,?PPI)。但由于PPI廣泛存在及其生理重要性,因此發(fā)現(xiàn)和確認(rèn)PPI藥理學(xué)靶點(diǎn)就非常具有吸引力。目前雖有一些文獻(xiàn)報(bào)道了PPI小分子抑制劑的例子,但僅有少數(shù)具有臨床意義。不能有效鑒別這類靶點(diǎn)主要?dú)w因于PPI相互作用界面的本質(zhì)特點(diǎn)。蛋白-小分子作用的接觸面大約為300~1000?2左右,而蛋白-蛋白相互作用的接觸面則為1500~3000?2之間。而且,?蛋白-蛋白相互接觸的表面往往是平坦的,不會(huì)存在很多溝槽或者口袋,?使得小分子化合物找不到適合的結(jié)合位點(diǎn)。盡管如此,但成功鑒別這些相互作用的藥物靶點(diǎn)對(duì)于人類健康事業(yè)極其重要。因此,近年來關(guān)于蛋白-蛋白相互作用小分子抑制劑的研究已經(jīng)成為全球藥物化學(xué)家關(guān)注的熱點(diǎn)。


表觀遺傳及組蛋白密碼

基因轉(zhuǎn)錄調(diào)控依賴于適當(dāng)?shù)霓D(zhuǎn)錄因子結(jié)合到靶基因啟動(dòng)子區(qū)域。盡管調(diào)控基因表達(dá)的信號(hào)復(fù)雜多樣,但其中一個(gè)主要的調(diào)控機(jī)制是通過DNA及組蛋白修飾來介導(dǎo)完成的。這些修飾標(biāo)記以及參與修飾和去除修飾的相關(guān)蛋白(酶),形成了轉(zhuǎn)錄調(diào)控的重要基礎(chǔ)。近年采用質(zhì)譜法鑒定出了上百種組蛋白翻譯后修飾(Posttranslational?modifications,?PTMs),其中最主要的PTMs存在于組蛋白的N-末端。雖然大多數(shù)修飾標(biāo)記的生理學(xué)意義還未知,但一些關(guān)鍵的翻譯后修飾已定義了明確的生物學(xué)活性。

可以說,到目前研究最深入的組蛋白翻譯后修飾就是賴氨酸乙?;?。組蛋白賴氨酸乙?;扇趸疍NA-組蛋白相互作用,募集相關(guān)轉(zhuǎn)錄因子。該修飾是基因進(jìn)行轉(zhuǎn)錄活化的標(biāo)志。乙酰賴氨酸(Acetyllysine)可被稱為溴結(jié)構(gòu)域的蛋白基序(motif)選擇性識(shí)別。目前在人類基因組中至少已鑒定出43種含溴結(jié)構(gòu)域的蛋白質(zhì)(如圖1),其中大多數(shù)為重要的轉(zhuǎn)錄調(diào)控因子。溴結(jié)構(gòu)域一般由60-120個(gè)氨基酸殘基組成。二級(jí)結(jié)構(gòu)為雙性α-螺旋,三級(jí)結(jié)構(gòu)為左手上下四螺旋束狀結(jié)構(gòu),含一個(gè)疏水穴(Hydrophobic?pocket)。已發(fā)現(xiàn)人類某些疾病與溴結(jié)構(gòu)域突變相關(guān)。


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表觀遺傳修飾閱讀器”reader”蛋白結(jié)構(gòu)域靶點(diǎn)的發(fā)現(xiàn)


??圖1.含溴結(jié)構(gòu)域蛋白聚類分析

盡管溴結(jié)構(gòu)域在生物醫(yī)學(xué)研究上獲得重大關(guān)注,但其他閱讀器”Reader”蛋白結(jié)構(gòu)域異常也具有顯著的生理學(xué)效應(yīng)?;蜣D(zhuǎn)座使得甲基賴氨酸閱讀器”reader”結(jié)構(gòu)域從JARID1A或PHF23與Nucleoporin-98融合,導(dǎo)致維持細(xì)胞多能性的基因過量表達(dá),而引起腫瘤發(fā)生。甲基賴氨酸閱讀器蛋白結(jié)構(gòu)域家族至少含有200個(gè)成員,分布于不同的亞家族。該大家族蛋白成員結(jié)構(gòu)域各異,與甲基化賴氨酸的位置和甲基化狀態(tài)具有高度特異性。與賴氨酸乙?;啾?組蛋白甲基化的發(fā)生具有位點(diǎn)特異性。同樣,許多甲基賴氨酸閱讀器“Reader”也是與被修飾殘基周圍的氨基酸殘基反應(yīng)(其提供了序列特異性結(jié)構(gòu)域)。這與賴氨酸甲基化在募集轉(zhuǎn)錄因子和其他染色質(zhì)修飾酶等方面的作用密切相關(guān)。此外,其他的組蛋白翻譯后修飾識(shí)別結(jié)構(gòu)域也被廣泛研究,包括結(jié)合磷酸化絲氨酸的BRCT家族、甲基化DNA結(jié)合蛋白(如MBD結(jié)構(gòu)域)、甲基精氨酸結(jié)合蛋白(如ADD結(jié)構(gòu)域)。一些閱讀器“reader”結(jié)構(gòu)域突變與某些疾病相關(guān)聯(lián),比如甲基CpG結(jié)合蛋白2(MeCP2)突變會(huì)破壞DNA結(jié)合基序(motif),會(huì)導(dǎo)致一種稱為Rett綜合癥的X染色體連鎖精神和生理發(fā)育遲滯癥。BRCT結(jié)構(gòu)域,識(shí)別磷酸化的組蛋白尾(histone?tails),參與到DNA損傷應(yīng)答元件的募集,BRCA1蛋白的BRCT結(jié)構(gòu)域發(fā)生突變,會(huì)增加家族性患乳腺癌和卵巢癌的風(fēng)險(xiǎn)。
?表觀遺傳修飾閱讀器”reader”蛋白結(jié)構(gòu)域靶點(diǎn)的發(fā)現(xiàn)

? 圖2.?甲基賴氨酸閱讀器結(jié)構(gòu)域具結(jié)構(gòu)多樣性

表觀遺傳與藥物發(fā)現(xiàn)
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由于表觀遺傳修飾閱讀器結(jié)構(gòu)域的生理重要性,那么藥理操縱這些蛋白相互作用(PPI)也為疾病管理提供了新的途徑。最先進(jìn)的一類以表觀遺傳閱讀器結(jié)構(gòu)域?yàn)榘悬c(diǎn)的抑制劑,來源于鑒定可提高肝細(xì)胞產(chǎn)生載脂蛋白A-1的分子表型篩選(phenotypic?screens)。機(jī)理研究從篩選中鑒定出了先導(dǎo)化合物以及相關(guān)的化合物(+)-JQ1,?作為BET家族溴結(jié)構(gòu)域的抑制劑。盡管ApoA-1水平升高對(duì)于心血管的好處仍存爭(zhēng)議,但是BET溴結(jié)構(gòu)域抑制已被證明能有效減緩NUT?midline?carubinas腫瘤(是一種罕見又致命的惡性皮膚癌,通常NMC患者的存活時(shí)間不足1年)生長。這一初始工作也有助于開發(fā)一種抗Midline?carcinomas(NCT01587703)腫瘤的新型化療藥物,目前已經(jīng)進(jìn)入臨床I期。另外一種BET溴結(jié)構(gòu)域抑制劑,RVX-208,也作為糖尿病前期葡萄糖代謝藥物進(jìn)入臨床II期。

由于BET家族溴結(jié)構(gòu)域抑制劑的偶然發(fā)現(xiàn),眾多學(xué)術(shù)及企業(yè)界研究團(tuán)體備受鼓舞,積極投身其他溴結(jié)構(gòu)域靶點(diǎn)的發(fā)現(xiàn)。一個(gè)結(jié)構(gòu)基因組學(xué)聯(lián)盟項(xiàng)目就已經(jīng)結(jié)晶解析出了很多溴結(jié)構(gòu)域,并針對(duì)不同溴結(jié)構(gòu)域篩選了選擇性的小分子抑制劑(
www.thesgc.org)?[注:Cayman公司可提供很多此類小分子化合物]。其他閱讀器結(jié)構(gòu)域的小分子抑制劑也被鑒定出來,如以L3MBTL和L3MBTL3的甲基賴氨酸結(jié)合結(jié)構(gòu)域?yàn)榘悬c(diǎn)小分子(如?UNC1215,?Cayman貨號(hào)#?13968)。

鑒于表觀遺傳修飾閱讀器結(jié)構(gòu)域在基因表達(dá)調(diào)控方面的重要性,那么這些蛋白基序家族則為藥物研發(fā)提供了新的有前景的領(lǐng)域。實(shí)際上,目前研究發(fā)現(xiàn)BET溴結(jié)構(gòu)域抑制劑不僅具有抗增值作用,還包括抗體病毒和抗炎癥的作用。為了幫助全球從事該領(lǐng)域的科研人員更好的解析決讀器蛋白結(jié)構(gòu)域,Cayman公司可提供多種純化的閱讀器結(jié)構(gòu)域、抑制劑篩選試劑盒以及化合物探針。隨著該領(lǐng)域的深入研究,也希望有更多新型的表觀修飾閱讀器結(jié)構(gòu)域抑制劑被鑒定出來,以此推動(dòng)藥物早期篩選的進(jìn)程。

------------------------------------------------------------------------- 文: 欣博盛市場(chǎng)部 Harry 編譯?-------------------------------------------------------------------------

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克羅莫結(jié)構(gòu)域及異染色質(zhì)蛋白(chromodomains?and?heterochromatin?proteins)?-- 結(jié)合mono,?di-tri-methyl?H3K4/9/27
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?14769?CBX2?chromodomain(human?recombinant)?Histone?H3K9me3?or?H3K27me3?≥90%
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?H3K9me2?and?H3K9me3,with?a?pre-
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??????????????????
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?recombinant)
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?gene?in?neurological?disorders,?such?as?
?schizophrenia?and?bipolar?affective?disorder.
?≥95%
?11071?BRD2?bromodomain1?(human?
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?A?member?of?the?BET?protein?family,?which?
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?histone?tails,?coupling?histone?acetylation?
?marks?to?the?transcriptional?regulation?of?target?
?promoters.
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?See?Item?No.?11068?for?details.?≥95%
?14822?BRD4?bromodomains?1?and?2?(human?
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?See?Item?No.?11068?for?details.?≥70%
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?component?of?the?switch/sucrose?nonfermen-
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?tumor?suppressor?role?by?acting?as?a?cofactor?
?for?p53?and?regulating?breast?cancer?tumori-
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?≥90%
?11509?BRD9?bromodomain?(human?
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?≥95%
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?acetylated?histone?H4,?Loss?of?BRDT?leads?to?
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?≥95%
?11649?BRDT?bromodomain?2?(human?
?recombinant)
?See?Item?No.?11548?for?details.?≥95%
?14492?BRDT?bromodomains?1?and?2?(human?
?recombinant)
?See?Item?No.?11548?for?details.?≥95%
?11284?BRG1?bromodomain?(human?
?recombinant)
?BRG1?is?a?member?of?the?SWI/SNF?proteins?
?family,?which?forms?part?of?the?large?ATP-
?dependent?chromatin?remodeling?complex?that?
?is?requires?for?transciptional?activation?of?genes?
?normally?repressed?by?chromatin.
?≥95%
?11289?BRM?bromodomain?(human?
?recombinant)
?BRM?is?a?member?of?the?SWI/SNF?proteins?
?family,?which?forms?part?of?the?large?ATP-
?dependent?chromatin?remodeling?complex?that?
?is?requires?for?transciptional?activation?of?genes?
?normally?repressed?by?chromatin.
?≥95%
?11650?BRPF3?bromodomain?1?(human?
?recombinant)
?BRPF3?is?a?component?of?the?MOZ/MORF?
?histone?acetytransferase(HAT)?complex.The?
?addition?of?BRPF?proteins?to?MOZ/MORF?
?increases?its?HAT?activity.This?product?contains?
?the?first?bromodomain?of?BRPF3.
?≥95%
?14133?CECR2?bromodomain?(human?
?recombinant)
?CECR2?is?a?transcription?factor?that?forms?a?
?heterodimeric?complex?with?the?ATP-dependent?
?chromation?remodeler?SNF2L?forming?the?
?CERF,?which?plays?a?critical?role?in?neurula-
?tion.?The?bromodomain?of?CECR2?has?strong?
?γ-H2AX?inhibition?activity?suggesting?that?CEC-
?R2?may?play?a?role?in?DNA?damage?response.
?≥95%
11288?CREB-binding?protein?bromodomain?(human?
?recombinant)
?The?CREBBP?bromodomain?has?been?shown?
?to?modulate?the?stability?and?function?of?the?
?tumor?suppressor?protein?p53.?CREBBP?brom-
?odomain?recognizes?the?acetylated?lysine?
?residue?382?on?p53.
?≥95%
11920?PCAF?bromodomain?(human?recombinant)?p300/CBP-associated?factor(PCAF)?is?a?tran-
?scriptional?coactivator?that?works?both?as?a?
?histone?lysine?acetytransferase,?through?its?
?HAT?domain,?and?as?an?acety-lysine?reader?
?throung?its?conserved?bromodomain?located?
?directly?C-terminal?to?the?HAT?domain.The?
?PCAF?bromodomain?binds?acetylated?histone?
?H3?and?H4?as?well?as?non-histone?targets.
?≥95%
11652?Polybromo-1D?bromodomain?1?(human?
?recombinant)
?PBRM1?contains?six?bromodomains?and?is?a?
?component?of?the?SWI/SNF?complex,?PBAF.?
?PBAF?is?target?to?acetylated?sites?in?chromatin?
?by?the?PBRM1?bromodomains,?where?it?plays?a?
?role?in?cell?cycle?regulation?and?tumor?
?suppression.
?≥95%
14659?SP140?PHD?and?bromodomain?(human?
?recombinant)
?The?C-terminal?PHD?and?bromodomain?
?regions?of?SP140?work?in?concert?to?bind?to?
?chromatin?and?regulate?gene?transcription.?
?Nuclear?bodies,?which?are?involved?in?the?
?pathogenesis?of?acute?promyelocytic?leukemia?
?and?viral?infection,?consist?of?several?
?components,such?as?Sp100,?NDP52,PML,?and?
?SP140.
?≥95%
11922?TAF1?bromodomain?1?(human?recombinant)?TAF1?is?a?component?of?transcription?factor?IID,?
?and?binds?to?core?promoter?sequences?at?the?
?transcription?start?site.?TAF1?helps?control?
?transcription?by?both?its?kinase?and?histone?
?acetyltranferase?enzymatic?activities.?This?
?protein?product?contains?the?first?bromodomain?
?of?TAF1.
?≥95%
14495?TAF1?bromodomain?2?(human?recombinant)?Contains?the?second?bromodomain?of?TAF1?≥95%
14494?TAF1?bromodomains?1?and?2?(human?
?recombinant)
?Contains?the?first?and?second?bromodomains?
?of?TAF1
?≥90%
11653?TRIM24?bromodomain?(human?recombinant)??TRIM24?is?a?transcriptional?confactor?whose?
?inactivation?leads?to?hepatocellular?carcinoma?
?in?mice.?The?N-terminal?TRIM?domain?of?TRIM-
?24?binds?ligand-bound?nuclear?receptors,?while?
?its?tandem?C-terminal?plant?homeo-domain?
?and?bromodomain?target?TRIM24?to?acetylated?
?histones?in?chromatin.
?≥95%
14660?TRIM28?PHD?and?bromodomain?(human?
?recombinant)?
?TRIM28,?a??KRAB?associated?protein,?in-
?teracts?with?SETD1?and?other?methyltrans-
?ferases?for?gene?silencing?of?endogenous?
?retroviruses?during?embryogenesis.The?PHD?
?finger?of?TRIM28?plays?an?important?role?in ?
?gene?silencing.The?PHD?domain?and?bromo-
?domain?of?TRIM28?work?together?to?facilitate?
?lysine?SUMOylation,?which?is?a?requirement?
?for?TRIM28?activity?in?gene?silencing.
?≥85%
14661?TRIM33?PHD?and?bromodomain?(human?
?recombinant)?
?TRIM33?is?a?multi-domain?regulator?of?
?transcription?that?is?necessary?for?emb-
?ryogenesis.?TRIM33?is?targeted?to?DNA?by?
?its?tandem?PHD?and?bromodomain,?which?
?bind?histone?3?at?H3K9me3?and?
?H3K18ac,respectively.
?≥80%
11549?WDR9?bromodomain?2?(human?recombinant)?WDR9?pessesses?two?bromodomain?
?motifs?and?eight?WD?repeats.It?is?also?
?known?to?interact?with?BRG1?(SMARCA4).
?≥85%


???????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????????
DNA甲基修飾閱讀器?(Readers)
?編號(hào)?產(chǎn)品名稱?識(shí)別位點(diǎn)?純度
?11286?MBD2?(human?recombinant;?methyl?binding?
?domain?aa?150-220
?5-Methylcytosine?in?promoters?on?
?CpG?islands
?≥95%
?11287?MeCP2?(human?recombinant;?
?methyl?binding?domain?aa?77-166
?5-Methylcytosine?in?promoters?on?
?CpG?islands;?high?affinity?binding?
?is?facilit-ated?by?DNA?fragments?
?containing?A/T?bases[A/T]≥4)?adj-
?acent?to?the?methyl-CpG.
?≥95%

溴結(jié)構(gòu)域TR-FRET分析試劑盒/Bromodomain?TR-FRET?Assay?Kits
--- 可用于高通量快速鑒定溴結(jié)構(gòu)域/乙?;南嗷プ饔玫囊种苿?/span>

表觀遺傳修飾閱讀器”reader”蛋白結(jié)構(gòu)域靶點(diǎn)的發(fā)現(xiàn)?
圖.?溴結(jié)構(gòu)域TR-FRET(時(shí)間分辨率熒光共振能量轉(zhuǎn)移)分析試劑盒原理示意圖。該檢測(cè)利用銪螯合物標(biāo)記的重組溴結(jié)構(gòu)域作為供體,乙?;慕M蛋白肽與鏈親和素-APC偶聯(lián)作為受體。銪螯合物受激發(fā)后釋放光子或傳遞給APC分子
??
?編號(hào)
?產(chǎn)品名稱
?600710
?BAZ2B?bromodomain?TR-FRET?Assay?Kit
?600500
?BRD2?bromodomain?1?TR-FRET?Assay?Kit
?600510
?BRD2?bromodomain?2?TR-FRET?Assay?Kit
?600810
?BRD2?bromodomains?1?and?2?TR-FRET?Assay?Kit
?600630
?BRD3?bromodomain?1?TR-FRET?Assay?Kit
?600820
?BRD3?bromodomains?1?and?2?TR-FRET?Assay?Kit
?600640
?BRD3?bromodomain?2?TR-FRET?Assay?Kit
?600520
?BRD4?bromodomain?1?TR-FRET?Assay?Kit
?600530
?BRD4?bromodomain?2?TR-FRET?Assay?Kit
?600830
?BRD4?bromodomains?1?and?2?TR-FRET?Assay?Kit
?600650
?BRDT?bromodomain?1?TR-FRET?Assay?Kit
?600720
?BRG1?bromodomain?TR-FRET?Assay?Kit
?600730
?BRM?bromodomain?TR-FRET?Assay?Kit
?600850
?CBP?bromodomain?TR-FRET?Assay?Kit
?600870
?TAF-1?bromodomain?1?TR-FRET?Assay?Kit
?600930
?TAF-1?bromodomains?1?and?2?TR-FRET?Assay?Kit


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