Hossain Md Shahadat, Alam Md Mahbubul, Huang Zhiwei, Mousazadeh Faeze, Sarangi Ronit, de Jong Ebbing, Kolamunna Kavindu C, Adhya Albert L, Hougland James L, Acharya Atanu, Mozhdehi Davoud
Department of Chemistry, Syracuse University, 111 College Place, Syracuse, New York 13244, United States.
Upstate Medical University, Proteomics and Mass Spectrometry, Weiskotten Hall 4307 WHA, 766 Irving Avenue, Syracuse, New York 13210, United States.
Bioconjug Chem. 2025 Mar 19;36(3):415-423. doi: 10.1021/acs.bioconjchem.4c00493. Epub 2025 Mar 3.
Geranylgeranylation is a critical post-translational modification essential for various cellular functions. However, current methods for synthesizing geranylgeranylated proteins are complex and costly, which hinders access to these proteins for both biophysical and biomaterials applications. Here, we present a method for the one-pot production of geranylgeranylated proteins in . We engineered to express geranylgeranyl pyrophosphate synthase (GGS), an enzyme that catalyzes the production of geranylgeranyl pyrophosphate. By coexpressing GGS with a geranylgeranyltransferase, we achieved efficient geranylgeranylation of model protein substrates, including intrinsically disordered elastin-like polypeptides (ELPs) and globular proteins such as mCherry and the small GTPases RhoA and Rap1B. We examined the biophysical behavior of the resulting geranylgeranylated proteins and observed that this modification affects the phase-separation and nanoassembly of ELPs and lipid bilayer engagement of mCherry. Taken together, our method offers a scalable, versatile, and cost-effective strategy for producing geranylgeranylated proteins, paving the way for advances in biochemical research, therapeutic development, and biomaterial engineering.
香叶基香叶基化是一种对多种细胞功能至关重要的翻译后修饰。然而，目前合成香叶基香叶基化蛋白的方法复杂且成本高昂，这阻碍了在生物物理和生物材料应用中获取这些蛋白。在此，我们展示了一种在[具体体系未给出]中一锅法生产香叶基香叶基化蛋白的方法。我们对[具体对象未给出]进行工程改造以表达香叶基香叶基焦磷酸合酶（GGS），该酶催化香叶基香叶基焦磷酸的产生。通过将GGS与香叶基香叶基转移酶共表达，我们实现了对模型蛋白底物的高效香叶基香叶基化，包括内在无序的弹性蛋白样多肽（ELP）以及球状蛋白，如mCherry和小GTP酶RhoA和Rap1B。我们研究了所得香叶基香叶基化蛋白的生物物理行为，并观察到这种修饰影响了ELP的相分离和纳米组装以及mCherry与脂质双层的结合。综上所述，我们的方法为生产香叶基香叶基化蛋白提供了一种可扩展、通用且具有成本效益的策略，为生物化学研究、治疗开发和生物材料工程的进展铺平了道路。
