Cloned (Comment) | Organism |
---|---|
recombinant expression of wild-type and mutant MdL1 and MdL2 in Pichia pastoris | Musca domestica |
Crystallization (Comment) | Organism |
---|---|
purified recombinant MdL2, sitting drop vapour diffusion method, 18°C, precipitant solution containing 28% isopropanol, 21% PEG 4000, and 0.115 M sodium citrate pH 4.2, addition of 0.2 M ammonium acetate, 30% 2-methyl-2,2-pentanediol, 0.1 M sodium citrate, pH 5.6, X-ray diffraction structure determination and analysis at 1.9 A resolution, modeling | Musca domestica |
Protein Variants | Comment | Organism |
---|---|---|
additional information | introduction of six basic residues onMdL1 surface increases by 1 unit the pH optimum for the activity upon bacterial walls | Musca domestica |
N46D/S106V/T107A | site-directed mutagenesis of isozyme MdL2, the mutant pKas are increased compared to the wild-type pKas | Musca domestica |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Musca domestica | - |
digestive lysozymes 1 and 2, i.e. MdL1 and MdL2 | - |
Purification (Comment) | Organism |
---|---|
recombinant wild-type and mutant MdL2s from Pichia pastoris by ammonium sulfate fractionation, and cation exchange, for the sextuple mutant, or anion exchange, for the other mutants, chromatography | Musca domestica |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
4-methylumbelliferyl-beta-D-N,N',N''-triacetylchitotrioside + H2O | - |
Musca domestica | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
More | structure analysis of MdL1 and MdL2, overview. Residues S106-T107 delimit a polar pocket around E32, as catalytic acid/base, and N46 contributes to the positioning of D50, as catalytic nucleophile | Musca domestica |
Synonyms | Comment | Organism |
---|---|---|
Mdl1 | - |
Musca domestica |
MdL2 | - |
Musca domestica |
pH Optimum Minimum | pH Optimum Maximum | Comment | Organism |
---|---|---|---|
additional information | - |
the acidic pH optimum for MdL2 and MdL1 activities upon methylumbelliferylchitotrioside is determined by the presence of N46, S106 and T107 in the environment of their catalytic residues, which favors pKas reduction. The acidic pH optimum upon bacterial walls is determined by a low concentration of positive charges on the MdL2 and MdL1 surfaces | Musca domestica |
General Information | Comment | Organism |
---|---|---|
additional information | comparison of the structure, surface charge, dissociation constants, and pH optimum of the fly enzyme with the enzyme from egg white, overview | Musca domestica |