Chemical Synthesis of the 20 kDa Heme Protein Nitrophorin 4 by α‐Ketoacid‐Hydroxylamine (KAHA) Ligation. Issue 44 (8th September 2015)

• Record Type: Journal Article
• Authors: He, Chunmao; Kulkarni, Sameer S.; Thuaud, Frédéric; Bode, Jeffrey W.
• Is Part Of: Angewandte Chemie international edition. Volume 54:Issue 44(2015)
• Page Start: 12996
• Page End: 13001
• Publication Date: 2015-09-08
• Subjects: heme proteins -- ligation reactions -- peptides -- protein synthesis -- solid‐phase peptide synthesis; Chemistry -- Periodicals; 540
• DOI: 10.1002/anie.201505379
• Languages: English
• ISSNs: 1433-7851

Abstract

Abstract: The chemical synthesis of the 184‐residue ferric heme‐binding protein nitrophorin 4 was accomplished by sequential couplings of five unprotected peptide segments using α‐ketoacid‐hydroxylamine (KAHA) ligation reactions. The fully assembled protein was folded to its native structure and coordinated to the ferric heme b cofactor. The synthetic holoprotein, despite four homoserine residues at the ligation sites, showed identical properties to the wild‐type protein in nitric oxide binding and nitrite dismutase reactivity. This work establishes the KAHA ligation as a valuable and viable approach for the chemical synthesis of proteins up to 20 kDa and demonstrates that it is well‐suited for the preparation of hydrophobic protein targets. Abstract : It′s all coming together! The 20 kDa heme protein nitrophorin 4 was synthesized by sequential KAHA ligations using a convergent strategy. The folded synthetic protein readily coordinates the heme cofactor, and the holoprotein shows identical biological properties to the wild‐type protein for NO binding and nitrite dismutase reactivity.