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	additional information	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	additional information	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	additional information	CerS6 knockdown by shRNA	753223	
2.3.1.24	additional information	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	additional information	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	additional information	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	additional information	enzyme knockout by shRNA	736471	
2.3.1.24	additional information	in HEK-293 cells, ectopically expressed murine CerS5 only sensitized cells to doxorubicin and vincristine	752796	
2.3.1.24	additional information	knockdown of CerS6 by siRNA. Profiles of non-hydroxylated and 2-hydroxy-ceramides in transgenic HeLa cells expressing different CerS isozymes and or different specific interfence RNAs, overview	754361	
2.3.1.24	additional information	LASS5 gene silencing by siRNAexogenously expressed LASS5 in lung epithelia is membrane-associated, triggering increased ceramide synthesis. Maximal inhibition is achieved when LASS5 is coexpressed with a plasmid encoding a neutral sphingomyelinase involved in sphingomyelin hydrolysis. Compared with control cells, cells transfected with LASS5 siRNA exhibit a 43% increase in rates of phosphatidylcholine synthesis	-, 754360	
2.3.1.24	additional information	no loss of activity is observed for the unglycosylated mutant (HA-tagged Lass6-N18Q) when compared with either the glycosylated mutant HA-tagged Lass6-N285Q or the glycosylated wild-type Lass6	752700	
2.3.1.24	additional information	profiles of non-hydroxylated and 2-hydroxy-ceramides in transgenic HeLa cells expressing different CerS isozymes and or different specific interfence RNAs, overview	754361	
2.3.1.24	N18Q	site-directed mutagenesis, a glycosylation site mutant	752700	
2.3.1.24	N285Q	site-directed mutagenesis, a putative glycosylation site mutant	752700	
2.3.1.24	S341A	mutation in potential phosphorylation site, about 45% of wild-type activity, substrate tetracosanoyl-CoA	754143	
2.3.1.24	S349A	mutation in potential phosphorylation site, about 90% of wild-type activity, substrate tetracosanoyl-CoA	754143	
2.3.1.24	T346A	mutation in potential phosphorylation site, about 80% of wild-type activity, substrate tetracosanoyl-CoA	754143