7REO
Crystal structure of an engineered variant of single-chain Penicillin G Acylase from Kluyvera cryocrescens (global hydrolysis Rd3CHis)
Summary for 7REO
Entry DOI | 10.2210/pdb7reo/pdb |
Descriptor | Penicillin G Acylase, CALCIUM ION (3 entities in total) |
Functional Keywords | penicillin, acylase, hydrolase |
Biological source | Kluyvera cryocrescens |
Total number of polymer chains | 1 |
Total formula weight | 86206.26 |
Authors | Orth, P. (deposition date: 2021-07-13, release date: 2021-11-17, Last modification date: 2023-10-18) |
Primary citation | Fryszkowska, A.,An, C.,Alvizo, O.,Banerjee, G.,Canada, K.A.,Cao, Y.,DeMong, D.,Devine, P.N.,Duan, D.,Elgart, D.M.,Farasat, I.,Gauthier, D.R.,Guidry, E.N.,Jia, X.,Kong, J.,Kruse, N.,Lexa, K.W.,Makarov, A.A.,Mann, B.F.,Milczek, E.M.,Mitchell, V.,Nazor, J.,Neri, C.,Orr, R.K.,Orth, P.,Phillips, E.M.,Riggins, J.N.,Schafer, W.A.,Silverman, S.M.,Strulson, C.A.,Subramanian, N.,Voladri, R.,Yang, H.,Yang, J.,Yi, X.,Zhang, X.,Zhong, W. A chemoenzymatic strategy for site-selective functionalization of native peptides and proteins. Science, 376:1321-1327, 2022 Cited by PubMed Abstract: 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. PubMed: 35709255DOI: 10.1126/science.abn2009 PDB entries with the same primary citation |
Experimental method | X-RAY DIFFRACTION (1.812 Å) |
Structure validation
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