化学
生物素
生物合成
生物化学
接口(物质)
脂肪酸
酶
肺表面活性物质
吉布斯等温线
作者
Aochiu Chen,Rebecca N. Re,Thomas B. Davis,Khai Tran,Yuta W. Moriuchi,Sitong Wu,James J. La Clair,G.V. Louie,M.E. Bowman,David J. Clarke,Logan Mackay,Dominic J. Campopiano,Joseph P. Noel,Michael D. Burkart
摘要
Site-specific covalent conjugation offers a powerful tool to identify and understand protein–protein interactions. In this study, we discover that sulfur fluoride exchange (SuFEx) warheads effectively crosslink the Escherichia coli acyl carrier protein (AcpP) with its partner BioF, a key pyridoxal 5′-phosphate (PLP)-dependent enzyme in the early steps of biotin biosynthesis by targeting a tyrosine residue proximal to the active site. We identify the site of crosslink by MS/MS analysis of the peptide originating from both partners. We further evaluate the BioF–AcpP interface through protein crystallography and mutational studies. Among the AcpP-interacting BioF surface residues, three critical arginine residues appear to be involved in AcpP recognition so that pimeloyl-AcpP can serve as the acyl donor for PLP-mediated catalysis. These findings validate an evolutionary gain-of-function for BioF, allowing the organism to build biotin directly from fatty acid biosynthesis through surface modifications selective for salt bridge formation with acidic AcpP residues.
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