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 | 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 variants 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 enzymes Vmax or Km, has no effect on its in vitro anti-pseudomonal activity, and does not reduce lytic function | Homo sapiens |
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 | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | - |
- |
- |
Source Tissue | Comment | Organism | Textmining |
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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 Maximum [°C] | Comment | Organism |
---|---|---|---|
25 | - |
assay at | Homo sapiens |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
7 | - |
assay at | Homo sapiens |
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 |