Hydrolytically Stable Maleimide-End-Functionalized Polymers for Site-Specific Protein Conjugation.
Hydrolytically Stable Maleimide-End-Functionalized Polymers for Site-Specific Protein Conjugation.
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DOI：
10.1021/acs.bioconjchem.1c00487
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发表时间：
2021-11-17
影响因子：
4.7
通讯作者：
Konkolewicz, Dominik
中科院分区：
文献类型：
--
作者：
Wright, Thaiesha A.;Rahman, Monica Sharfin;Bennett, Camaryn;Johnson, Madolynn R.;Fischesser, Henry;Ram, Natasha;Tyler, Amoni;Page, Richard C.;Konkolewicz, Dominik
蛋白质的半胱氨酸位点特异性偶联通常利用酯基团将马来酰亚胺或烯烃基团连接到聚合物上。然而，酯基容易水解，可能会失去通过生物偶联获得的好处。在这里，我们提出了一种简单的偶联策略，利用传统的1-乙基-3-（3-二甲氨基丙基）碳二亚胺/ n -羟基琥珀酰亚胺偶联的酰胺键稳定性，同时引入位点特异性。采用可逆加成-断裂链转移（RAFT）聚合技术合成了水解稳定的马来酰亚胺端功能化聚合物，可与蛋白质的游离半胱氨酸位点偶联。利用碳二亚胺为基础的化学方法，用呋喃保护的氨基乙基马来酰亚胺对聚合物的α端进行修饰。最后，通过反向diols - alder反应暴露马来酰亚胺，生成马来酰亚胺基团，从而实现巯基马来酰亚胺的生物偶联。利用EDC/NHS偶联、位点特异性羟乙基马来酰亚胺（HEMI）端功能化偶联、丙烯酸羟乙基（HEA）端功能化偶联和氨基乙基马来酰亚胺（AEMI）端功能化偶联等多种偶联策略，对结瘤铁杆菌（Fervidobacterium nodosum）中的一种嗜热纤维素酶（FnCel5a）进行偶联。只有EDC和AEMI偶联的聚合物在附着一周后仍保持偶联。这表明，水解稳定的酰胺基马来酰亚胺对于需要长期稳定性的偶联物来说是一种重要的生物偶联策略，而酯类更适合于需要随着时间的推移而脱键的聚合物系统。
Site-specific conjugation to cysteines of proteins often utilizes ester groups to link maleimide or alkene groups to polymers. However, the ester group is susceptible to hydrolysis, potentially losing the benefits gained through bioconjugation. Here we present a simple conjugation strategy that utilizes the amide bond stability of traditional 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide/N-hydroxysuccinimide coupling while introducing site-specificity. Hydrolytically stable maleimide-end functionalized polymers for site-specific conjugation to free cysteines of proteins were synthesized using reversible addition-fragmentation chain transfer (RAFT) polymerization. The polymers’ alpha terminus was amidated with a furan-protected aminoethyl maleimide using carbodiimide based chemistry Finally, the maleimide was exposed by a retro Diels-Alder reaction to yield the maleimide group allowing for thiol-maleimide click chemistry for bioconjugation. A thermophilic cellulase from Fervidobacterium nodosum (FnCel5a) was conjugated utilizing various strategies including random EDC/NHS coupling, site-specific hydroxyethyl maleimide (HEMI) end functionalized coupling, hydroxyethyl acrylate (HEA) end functionalized coupling, and amidoethyl maleimide (AEMI) end functionalized coupling. Only the polymers conjugated by EDC and AEMI remained conjugated a week after attachment. This indicates that hydrolytically stable amide based maleimides are an important bioconjugation strategy for conjugates that require long term stability, while esters are better suited to systems which require the debonding of the polymer over time.
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影响因子：
4.9
作者：
Falatach, Rebecca;McGlone, Cameron;Konkolewicz, Dominik
通讯作者：
Konkolewicz, Dominik
DOI：
10.1039/c8cc00782a
发表时间：
2018-03-25
期刊：
Chemical communications (Cambridge, England)
影响因子：
--
作者：
Sinha J;Podgórski M;Huang S;Bowman CN
通讯作者：
Bowman CN
影响因子：
4.2
作者：
Jurado, Encarnacion;Garcia-Roman, Miguel;Luis Jimenez-Perez, Jose
通讯作者：
Luis Jimenez-Perez, Jose
影响因子：
6.7
作者：
Kelestemur, Seda;Altunbek, Mine;Culha, Mustafa
通讯作者：
Culha, Mustafa
影响因子：
15
作者：
De, Priyadarsi;Li, Ming;Sumerlin, Brent S.
通讯作者：
Sumerlin, Brent S.