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Literature summary for 3.2.1.17 extracted from

  • Gill, A.; Scanlon, T.C.; Osipovitch, D.C.; Madden, D.R.; Griswold, K.E.
    Crystal structure of a charge engineered human lysozyme having enhanced bactericidal activity (2011), PLoS ONE, 6, e16788.
    View publication on PubMedView publication on EuropePMC

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant mutant R101D/R115H, 7 mg/ml protein in 10 mM potassium phosphate, pH 6.0, 100 mM NaCl, is mixed with crystallization solution containing 20 mM sodium acetate, pH 4.3, and 1.25 M NaCl, 18°C, 3-4 days, X-ray diffraction structure determination and analysis at 2.04 A resolution, molecular replacement and modeling Homo sapiens

Protein Variants

Protein Variants Comment Organism
R101D/R115H the charge engineered variant's two mutated amino acids exhibit stabilizing interactions with adjacent native residues, the mutant shows severalfold increased activity compared to the wild-type enzyme, the mutations cause no gross structural perturbations or loss of stability, but dramatically expand the negative electrostatic potential that, in the wild-type enzyme, is restricted to a small region near the catalytic residues. Reduction in the overall strength of the engineered enzyme's electrostatic potential field, the specific nature of this remodeled field underlies the variant’s reduced susceptibility to inhibition by anionic biopolymers, overview. The double mutant lyses bacteria effectively at alginate, mucin and DNA concentrations that inactivate wild-type enzyme. The mutations does not substantially impair the enzyme’s Vmax or Km, has no effect on its in vitro anti-pseudomonal activity, and does not reduce lytic function Homo sapiens

Inhibitors

Inhibitors Comment Organism Structure
alginate inactivation of the wild-type enzyme at high concentrations Homo sapiens
DNA inactivation of the wild-type enzyme at high concentrations Homo sapiens
F-actin inhibition of the wild-type enzyme Homo sapiens
mucin inactivation of the wild-type enzyme at high concentrations Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
-
-

Source Tissue

Source Tissue Comment Organism Textmining

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information lysis of Micrococcus luteus bacteria, the double mutant lyses bacteria effectively at alginate, mucin and DNA concentrations that inactivate wild-type enzyme Homo sapiens ?
-
?

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Homo sapiens

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Homo sapiens

General Information

General Information Comment Organism
physiological function human lysozyme is a key component of the innate immune system. But the wild type protein 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 possesses improved antibacterial activity in the presence of disease associated inhibitory molecules Homo sapiens