EC Number   |
Protein Variants |
Reference |
|---|
    2.3.1.24 | H212A |
site-directed mutagenesis |
736471 |
| 2.3.1.24 | H215D |
site-directed mutagenesis, the point mutation, changing a highly conserved histidine 215 into glutamate, in the Lag1 motif, which inhibits ceramide synthase function in the Lass1 and 5. With schlankH215D, an increase in ceramide levels cannot be observed |
753417 |
| 2.3.1.24 | H220A/H221A |
site-directed mutagenesis, mutation of the two residues involved in catalytic activity completely abrogates CerS5 activity in a constitutive dimer |
754101 |
| 2.3.1.24 | more |
a mutant based on the backbone of CerS4, containing an 11-residue sequence of a loop located between the last two putative transmembrane domainsfrom CerS2 (which generates C22-C24-ceramides), is altered such that it displays significant activity toward C24:1-CoA |
757217 |
| 2.3.1.24 | more |
a mutant based on the backbone of CerS5 (which generates C16-ceramide), containing an 11-residue sequence of a loop located between the last two putative transmembrane domainsfrom CerS2 (which generates C22-C24-ceramides), is altered such that it generates C22-C24 and other ceramides. The mutant generates C22:0-ceramide and C16:0-ceramide to a similar extent but also generates C18:0-, C20:0-, and C24:1-ceramides |
757217 |
| 2.3.1.24 | more |
CerS6 knockdown by shRNA |
753223 |
| 2.3.1.24 | more |
cloning of a chimeric HA-tagged CerS5:CerS2 isozymes heterodimer, insertion of a transmembrane (TM) domain3 between the two monomers of the dimer (CerS5:TM:CerS5-HA). The truncated mutant DELTA332-392 lacking the last putative transmembrane domain is inactive. Chimeric mutant CerS5:TM:CerS2-HA displays slightly more activity using C16-CoA as substrate than CerS5, but remarkably, CerS2 activity measured using C22-CoA is elevated by 3fold. Isozymes CerS5 and CerS6 modulate CerS2 activity upon coexpression. This increase in CerS2 activity is abolished using a noncatalytically active form of CerS5 in the constitutive dimer (CerS5HH:TM:CerS2-HA), demonstrating that optimal CerS2 activity depends on an interaction with a catalytically active form of CerS5 |
754101 |
| 2.3.1.24 | more |
complementation of the growth defect of DELTAlag1/DELTAlac1 yeast deletion mutant by recombinant GST-/FLAG-tagged LOH2. Complementation with LOH2 leads to the accumulation of C16-containing inositolphosphoceramides and ceramides indicating this isoform's preference for C16 fatty acids. Microsomes isolated from LOH2 overexpressing plants showed high levels of C16-ceramide synthase activity compared to microsomes from wild-type plants, indicating that LOH2 overexpression results in accumulation of functional enzyme |
752718 |
| 2.3.1.24 | more |
construction of larvae carrying transgenic UASschlankRNAi or UASschlankHA in combination with the hsGAL4 driver line. A short heat shock (1 h) induces schlank RNAi knockdown or schlank overexpression. Modulation of schlank activity correlates with the rate of ceramide de novo synthesis |
753417 |
| 2.3.1.24 | more |
enzyme knockout by shRNA |
736471 |
