| 2004 |
Human FVT-1 (KDSR) functions as a 3-ketodihydrosphingosine (KDS) reductase: recombinant hFVT-1 exhibits NADPH-dependent KDS reductase activity in vitro, and forced expression in TSC10-null yeast suppresses growth defects, establishing it as the mammalian KDS reductase. |
In vitro enzyme assay with purified recombinant protein, yeast complementation (TSC10-null rescue), overexpression in cultured cells |
The Journal of biological chemistry |
High |
15328338
|
| 2004 |
hFVT-1 (KDSR) localizes to the endoplasmic reticulum, and the large hydrophilic domain containing putative active-site residues faces the cytosolic side of the ER membrane, indicating that KDS is reduced to dihydrosphingosine on the cytosolic face of the ER. |
Immunofluorescence microscopy (ER colocalization) and proteinase K digestion topology assay |
The Journal of biological chemistry |
High |
15328338
|
| 2009 |
FVT1 (KDSR) is the principal 3-ketosphinganine reductase in mammalian cells: siRNA silencing of FVT1 directly reduced cellular reductase activity in proportion to FVT1 levels. The N-terminal membrane-spanning domain of FVT1 (absent in yeast Tsc10p) targets it to the ER lumen and confers distinct topology relative to yeast Tsc10p. Mutation of conserved catalytic residues differentially affected FVT1 vs. Tsc10p activity, revealing mechanistic differences between the two orthologs. |
siRNA knockdown with enzymatic activity measurement; N-terminal GFP fusion for ER-targeting domain mapping; factor Xa protease domain removal; active-site mutagenesis |
Journal of lipid research |
High |
19141869
|
| 2007 |
A missense mutation (Ala-175→Thr) in bovine FVT1 (KDSR) abolishes 3-ketodihydrosphingosine reductase activity in vitro, yet the mutant protein retains sufficient residual in vivo activity to complement a yeast knockout, explaining why SMA-affected calves are viable but develop neuron-specific degeneration. |
In vitro enzyme assay comparing Ala-175 vs. Thr-175 variants; yeast complementation assay |
Proceedings of the National Academy of Sciences of the United States of America |
High |
17420465
|
| 2017 |
Loss-of-function mutations in KDSR reduce ceramide levels in skin and impair KDSR enzymatic activity, causing defective acylceramide synthesis and leading to skin hyperkeratosis; thrombocytopenia is also present, indicating KDSR activity is required for normal platelet function. |
Whole-exome sequencing identifying mutations; KDSR enzymatic activity measurement in patient samples; ceramide level quantification in skin |
The Journal of investigative dermatology |
Medium |
28774589
|
| 2017 |
KDSR mutations causing exon skipping (including a recurrent silent third-base change) disrupt KDSR function as demonstrated by yeast complementation failure, establishing that loss of KDSR enzymatic activity underlies progressive symmetric erythrokeratoderma. |
Splicing assay (cDNA sequencing), yeast complementation, immunohistochemistry |
American journal of human genetics |
Medium |
28575652
|
| 2018 |
KDSR insufficiency impairs proplatelet formation from megakaryocytes: CD34+-derived megakaryocytes from KDSR-deficient patients showed hyperproliferation and reduced proplatelet formation, reversed by re-expression of functional KDSR in iPSC-derived megakaryocytes. Kdsr depletion in zebrafish recapitulated thrombocytopenia. A compensatory in vivo pathway partially normalizes downstream ceramide levels. |
CD34+ stem cell-derived megakaryocyte culture, iPSC differentiation with KDSR rescue, zebrafish kdsr morpholino knockdown, broad metabolomics screen |
Haematologica |
High |
30467204
|
| 2022 |
In patients with biallelic KDSR mutations, the KDSR substrate 3-ketodihydrosphingosine accumulates and is processed by ceramide synthases to produce novel keto-type ceramides (up to 10% of ceramide species) in the stratum corneum, revealing a bypass pathway when KDSR is non-functional and demonstrating that tight intermediate regulation during sphingolipid anabolism is required for normal ceramide composition. |
Stratum corneum lipid analysis by mass spectrometry in KDSR-mutant patients vs. controls; structural identification of keto-type ceramides |
Human molecular genetics |
Medium |
34686882
|