Scalable One-Pot Production of Geranylgeranylated Proteins in Engineered Prokaryotes DOI Creative Commons
Md Shahadat Hossain, Md. Mahbubul Alam, Zhiwei Huang

et al.

Bioconjugate Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 3, 2025

Geranylgeranylation is a critical post-translational modification essential for various cellular functions. However, current methods synthesizing geranylgeranylated proteins are complex and costly, which hinders access to these both biophysical biomaterials applications. Here, we present method the one-pot production of in Escherichia coli. We engineered E. coli express geranylgeranyl pyrophosphate synthase (GGS), an enzyme that catalyzes pyrophosphate. By coexpressing GGS with geranylgeranyltransferase, achieved efficient geranylgeranylation model protein substrates, including intrinsically disordered elastin-like polypeptides (ELPs) globular such as mCherry small GTPases RhoA Rap1B. examined behavior resulting observed this affects phase-separation nanoassembly ELPs lipid bilayer engagement mCherry. Taken together, our offers scalable, versatile, cost-effective strategy producing proteins, paving way advances biochemical research, therapeutic development, biomaterial engineering.

Language: Английский

Scalable One-Pot Production of Geranylgeranylated Proteins in Engineered Prokaryotes DOI Creative Commons
Md Shahadat Hossain, Md. Mahbubul Alam, Zhiwei Huang

et al.

Bioconjugate Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: March 3, 2025

Geranylgeranylation is a critical post-translational modification essential for various cellular functions. However, current methods synthesizing geranylgeranylated proteins are complex and costly, which hinders access to these both biophysical biomaterials applications. Here, we present method the one-pot production of in Escherichia coli. We engineered E. coli express geranylgeranyl pyrophosphate synthase (GGS), an enzyme that catalyzes pyrophosphate. By coexpressing GGS with geranylgeranyltransferase, achieved efficient geranylgeranylation model protein substrates, including intrinsically disordered elastin-like polypeptides (ELPs) globular such as mCherry small GTPases RhoA Rap1B. examined behavior resulting observed this affects phase-separation nanoassembly ELPs lipid bilayer engagement mCherry. Taken together, our offers scalable, versatile, cost-effective strategy producing proteins, paving way advances biochemical research, therapeutic development, biomaterial engineering.

Language: Английский

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