Application | Comment | Organism |
---|---|---|
drug development | rational redesign of EGC toward the synthesis of novel ganglioside-derived therapeutics | Rhodococcus sp. |
Cloned (Comment) | Organism |
---|---|
subcloned into pET28a using the NdeI/XhoI restriction sites and expressed in Escherichia coli BL21 | Rhodococcus sp. |
Crystallization (Comment) | Organism |
---|---|
by vapor diffusion method, crystals of the native enzyme belong to space group P21 with unit cell dimensions a=53.8 A, b=92.9 Å, c=94.5 A,beta=98.6°, to 1.6 A resolution. Crystal structure of mutant E351S in complex with the ganglioside GM3, crystals belong to space group C2, with unit cell dimensions a = 77.8 A, b = 62.0 A, c = 102.8 A, beta = 112.3°, to 1.1 A resolution. Active site of endo-glycoceramidase is split into a wide, polar cavity to bind the polyhydroxylated oligosaccharide moiety and a narrow, hydrophobic tunnel to bind the ceramide lipid chains | Rhodococcus sp. |
Protein Variants | Comment | Organism |
---|---|---|
E233A | glycosyl-enzyme intermediate trapped by soaking crystals of the general acid/base knock-out mutant E233A, with an activated sugar donor, 2,4-dinitrophenyl beta-D-lactoside | Rhodococcus sp. |
E351S | prevents substrate hydrolysis during crystallization | Rhodococcus sp. |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.04 | - |
D-erythro-sphingosine | - |
Rhodococcus sp. | |
0.3 | - |
D-erythro-dihydrosphingosine | - |
Rhodococcus sp. |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | - |
Rhodococcus sp. | 16020 | - |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Rhodococcus sp. | - |
- |
- |
Purification (Comment) | Organism |
---|---|
by Ni(II) affinity chromatography to more than 95% purity | Rhodococcus sp. |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
D-erythro-dihydrosphingosine + H2O | - |
Rhodococcus sp. | ? | - |
? | |
D-erythro-sphingosine + H2O | - |
Rhodococcus sp. | ? | - |
? | |
ganglioside + H2O | - |
Rhodococcus sp. | ? | - |
? |
Synonyms | Comment | Organism |
---|---|---|
EGC | - |
Rhodococcus sp. |
endo-glycoceramidase II | - |
Rhodococcus sp. |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.07 | - |
D-erythro-sphingosine | - |
Rhodococcus sp. | |
0.12 | - |
D-erythro-dihydrosphingosine | - |
Rhodococcus sp. |