A chemoenzymatic strategy for site-selective functionalization of native peptides and proteins

The emergence of new therapeutic modalities requires complementary tools for their efficient syntheses. Availability of methodologies for site-selective modification of biomolecules remains a long-standing challenge, given the inherent complexity and the presence of repeating residues that bear func...

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Published in:	Science (American Association for the Advancement of Science) 2022-06, Vol.376 (6599), p.1321-1327
Main Authors:	Fryszkowska, Anna, An, Chihui, Alvizo, Oscar, Banerjee, Goutami, Canada, Keith A., Cao, Yang, DeMong, Duane, Devine, Paul N., Duan, Da, Elgart, David M., Farasat, Iman, Gauthier, Donald R., Guidry, Erin N., Jia, Xiujuan, Kong, Jongrock, Kruse, Nikki, Lexa, Katrina W., Makarov, Alexey A., Mann, Benjamin F., Milczek, Erika M., Mitchell, Vesna, Nazor, Jovana, Neri, Claudia, Orr, Robert K., Orth, Peter, Phillips, Eric M., Riggins, James N., Schafer, Wes A., Silverman, Steven M., Strulson, Christopher A., Subramanian, Nandhitha, Voladri, Rama, Yang, Hao, Yang, Jie, Yi, Xiang, Zhang, Xiyun, Zhong, Wendy
Format:	Article
Language:	eng
Subjects:	Drug development Enzymatic activity Enzymes Functional groups Hormones Insulin Peptides Protecting groups Proteins Reagents Selectivity
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Summary:	The emergence of new therapeutic modalities requires complementary tools for their efficient syntheses. Availability of methodologies for site-selective modification of biomolecules remains a long-standing challenge, given the inherent complexity and the presence of repeating residues that bear functional groups with similar reactivity profiles. We describe a bioconjugation strategy for modification of native peptides relying on high site selectivity conveyed by enzymes. We engineered penicillin G acylases to distinguish among free amino moieties of insulin (two at amino termini and an internal lysine) and manipulate cleavable phenylacetamide groups in a programmable manner to form protected insulin derivatives. This enables selective and specific chemical ligation to synthesize homogeneous bioconjugates, improving yield and purity compared to the existing methods, and generally opens avenues in the functionalization of native proteins to access biological probes or drugs. Enzymes for tailored insulin derivatives The development of drugs based on protein hormones such as insulin is a challenge because many of the usual tools in organic synthesis are either not compatible with folded proteins or lack selectivity when redundant functional groups are present. Fryszkowska et al . generated a panel of enzymes that selectively modify one or more of the three amine groups of insulin, adding or removing protecting groups (see the Perspective by Lin and Chou). The enzymatically functionalized molecules can then be modified by electrophilic reagents to produce homogeneous insulin conjugates. The authors also demonstrate enzymatic activity on a range of bioactive peptides, suggesting that biocatalytic approaches may help in the development of potential peptide-based therapeutics or probes. —MAF Selective enzymatic modification of native proteins enables efficient synthesis of insulin therapeutics.
ISSN:	0036-8075 1095-9203