title: Crystal structure of a charge engineered human lysozyme having enhanced bactericidal activity.
abstract: Human lysozyme is a key component of the innate immune system, and recombinant forms of the enzyme represent promising leads in the search for therapeutic agents able to treat drug-resistant infections. The wild type protein, however, fails to participate effectively in clearance of certain infections due to inherent functional limitations. For example, wild type lysozymes are subject to electrostatic sequestration and inactivation by anionic biopolymers in the infected airway. A charge engineered variant of human lysozyme has recently been shown to possess improved antibacterial activity in the presence of disease associated inhibitory molecules. Here, the 2.04 Å crystal structure of this variant is presented along with an analysis that provides molecular level insights into the origins of the protein's enhanced performance. The charge engineered variant's two mutated amino acids exhibit stabilizing interactions with adjacent native residues, and from a global perspective, the mutations cause no gross structural perturbations or loss of stability. Importantly, the two substitutions dramatically expand the negative electrostatic potential that, in the wild type enzyme, is restricted to a small region near the catalytic residues. The net result is a reduction in the overall strength of the engineered enzyme's electrostatic potential field, and it appears that the specific nature of this remodeled field underlies the variant's reduced susceptibility to inhibition by anionic biopolymers.
authors.affiliation: Thayer School of Engineering, Dartmouth College, Hanover, New Hampshire, United States of America.
authors.forename: Avinash
authors.initials: A
authors.lastname: Gill
authors.forename: Thomas C
authors.initials: TC
authors.lastname: Scanlon
authors.forename: Daniel C
authors.initials: DC
authors.lastname: Osipovitch
authors.forename: Dean R
authors.initials: DR
authors.lastname: Madden
authors.forename: Karl E
authors.initials: KE
authors.lastname: Griswold
mesh_terms: D000199:Actins / Q000378:metabolism; D000900:Anti-Bacterial Agents / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D055162:Biocatalysis / Q000187:drug effects; D018360:Crystallography, X-Ray; D004795:Enzyme Stability / Q000187:drug effects; D006801:Humans; D008958:Models, Molecular; D009113:Muramidase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism* / Q000494:pharmacology; D050505:Mutant Proteins / Q000737:chemistry; D015202:Protein Engineering; D011499:Protein Processing, Post-Translational / Q000187:drug effects; D020539:Sequence Analysis, Protein; D055672:Static Electricity
chemical_list: D000199:Actins; D000900:Anti-Bacterial Agents; D050505:Mutant Proteins; D009113:Muramidase