title: Crystallographic snapshot of glycosylasparaginase precursor poised for autoprocessing.
abstract: Glycosylasparaginase belongs to a family of N-terminal nucleophile hydrolases that autoproteolytically generate their mature enzymes from single-chain protein precursors. Previously, based on a precursor structure paused at pre-autoproteolysis stage by a reversible inhibitor (glycine), we proposed a mechanism of intramolecular autoproteolysis. A key structural feature, a highly strained conformation at the scissile peptide bond, had been identified and was hypothesized to be critical for driving autoproteolysis through an N-O acyl shift. To examine this "twist-and-break" hypothesis, we report here a 1. 9-Å-resolution structure of an autoproteolysis-active precursor (a T152C mutant) that is free of inhibitor or ligand and is poised to undergo autoproteolysis. The current crystallographic study has provided direct evidence for the natural conformation of the glycosylasparaginase autocatalytic site without influence from any inhibitor or ligand. This finding has confirmed our previous proposal that conformational strain is an intrinsic feature of an active precursor.
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118-2526, USA.
authors.forename: Yeming
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Physiology and Biophysics, Boston University School of Medicine, 715 Albany Street, Boston, MA 02118-2526, USA. Electronic address: hcguo@bu.edu.
authors.forename: Hwai-Chen
authors.initials: HC
authors.lastname: Guo
mesh_terms: D001227:Aspartylglucosylaminase / Q000737:chemistry*; D001426:Bacterial Proteins / Q000737:chemistry*; D018360:Crystallography, X-Ray; D005417:Flavobacterium / Q000201:enzymology*; D008958:Models, Molecular; D050505:Mutant Proteins / Q000737:chemistry; D011498:Protein Precursors / Q000737:chemistry*; D017434:Protein Structure, Tertiary
chemical_list: D001426:Bacterial Proteins; D050505:Mutant Proteins; D011498:Protein Precursors; D001227:Aspartylglucosylaminase