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

被引：16

作者：

Fryszkowska, Anna ^{[1
]}

An, Chihui ^{[1
]}

Alvizo, Oscar ^{[2
]}

Banerjee, Goutami ^{[2
,4
]}

Canada, Keith A. ^{[1
]}

Cao, Yang ^{[1
]}

DeMong, Duane ^{[1
]}

Devine, Paul N. ^{[1
,5
]}

Duan, Da ^{[2
]}

Elgart, David M. ^{[2
]}

Farasat, Iman ^{[1
,6
]}

Gauthier, Donald R. ^{[1
]}

Guidry, Erin N. ^{[3
]}

Jia, Xiujuan ^{[1
]}

Kong, Jongrock ^{[1
]}

Kruse, Nikki ^{[2
]}

Lexa, Katrina W. ^{[3
,7
]}

Makarov, Alexey A. ^{[1
]}

Mann, Benjamin F. ^{[1
]}

Milczek, Erika M. ^{[1
,8
]}

Mitchell, Vesna ^{[2
]}

Nazor, Jovana ^{[2
]}

Neri, Claudia ^{[1
]}

Orr, Robert K. ^{[1
]}

Orth, Peter ^{[3
,9
]}

Phillips, Eric M. ^{[1
]}

Riggins, James N. ^{[2
]}

Schafer, Wes A. ^{[1
]}

Silverman, Steven M. ^{[1
]}

Strulson, Christopher A. ^{[1
]}

Subramanian, Nandhitha ^{[2
,10
]}

Voladri, Rama ^{[2
,11
]}

Yang, Hao ^{[1
]}

Yang, Jie ^{[2
,4
]}

Yi, Xiang ^{[2
,12
]}

Zhang, Xiyun ^{[2
,4
]}

Zhong, Wendy ^{[1
]}

机构：

[1] Merck & Co Inc, Proc Res & Dev, Rahway, NJ 07065 USA

[2] Codexis Inc, 200 Penobscot Dr, Redwood City, CA 94063 USA

[3] Merck & Co Inc, Discovery Chem, Kenilworth, NJ 07033 USA

[4] Fornia BioSolut Inc, 3876 Bay Ctr Pl, Hayward, CA 94545 USA

[5] Schrodinger Inc, 120 W 45th St,17th Floor, New York, NY 10036 USA

[6] Janssen Pharmaceut Co Johnson & Johnson, 1125 Trenton Harbourton Rd, Titusville, NJ 08560 USA

[7] Denali Therapeut Inc, 161 Oyster Point Blvd, San Francisco, CA 94080 USA

[8] Curie Co Inc, 6 Davis Dr, Durham, NC 27709 USA

[9] Proteovant Therapeut, 151 W 42nd St, New York, NY 10036 USA

[10] Lonza Biol Plc, Gonville Bldg B200,Chesterford Res Pk, Saffron Walden CB10 1XL, England

[11] Clorox Co, 4900 Johnson Dr, Pleasanton, CA 94080 USA

[12] Amgen Inc, 1120 Vet Blvd, San Francisco, CA 94080 USA

来源：

SCIENCE | 2022年 / 376卷 / 6599期

关键词：

PENICILLIN-G ACYLASE; SUBSTRATE-SPECIFICITY; AMINO-ACIDS; INSULIN; ACYLATION; RESIDUES; KINETICS; MUTANT;

D O I：

10.1126/science.abn2009

中图分类号：

O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];

学科分类号：

07 ; 0710 ; 09 ;

摘要：

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.

引用

页码：1321 / +

页数：163

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