The crystal structure of AtPDF1B [Arabidopsis thaliana PDF (peptide deformylase) 1B; EC 126.96.36.199], a plant specific deformylase, has been determined at a resolution of 2.4 Å (1 Å=0.1 nm). The overall fold of AtPDF1B is similar to other peptide deformylases that have been reported. Evidence from the crystal structure and gel filtration chromatography indicates that AtPDF1B exists as a symmetric dimer. PDF1B is essential in plants and has a preferred substrate specificity towards the PS II (photosystem II) D1 polypeptide. Comparative analysis of AtPDF1B, AtPDF1A, and the type 1B deformylase from Escherichia coli, identifies a number of differences in substrate binding subsites that might account for variations in sequence preference. A model of the N-terminal five amino acids from the D1 polypeptide bound in the active site of AtPDF1B suggests an influence of Tyr178 as a structural determinant for polypeptide substrate specificity through hydrogen bonding with Thr2 in the D1 sequence. Kinetic analyses using a polypeptide mimic of the D1 N-terminus was performed on AtPDF1B mutated at Tyr178 to alanine, phenylalanine or arginine (equivalent residue in AtPDF1A). The results suggest that, whereas Tyr178 can influence catalytic activity, other residues contribute to the overall preference for the D1 polypeptide.
Insights into the substrate specificity of plant peptide deformylase, an essential enzyme with potential for the development of novel biotechnology applications in agriculture
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Lynnette M. A. Dirk, Jack J. Schmidt, Yiying Cai, Jonathan C. Barnes, Katherine M. Hanger, Nihar R. Nayak, Mark A. Williams, Robert B. Grossman, Robert L. Houtz, David W. Rodgers; Insights into the substrate specificity of plant peptide deformylase, an essential enzyme with potential for the development of novel biotechnology applications in agriculture. Biochem J 1 August 2008; 413 (3): 417–427. doi: https://doi.org/10.1042/BJ20071641
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