[HTML][HTML] Quantification of 3-ketodihydrosphingosine using HPLC-ESI-MS/MS to study SPT activity in yeast Saccharomyces cerevisiae [S]

J Ren, J Snider, MV Airola, A Zhong, NA Rana… - Journal of Lipid …, 2018 - ASBMB
Serine palmitoyltransferase (SPT) catalyzes the rate-limiting step of condensation of L-
serine and palmitoyl-CoA to form 3-ketodihydrosphingosine (3KDS). Here, we report a
HPLC-ESI-MS/MS method to directly quantify 3KDS generated by SPT. With this technique,
we were able to detect 3KDS at a level comparable to that of dihydrosphingosine in yeast
Saccharomyces cerevisiae. An in vitro SPT assay measuring the incorporation of deuterated
serine into deuterated 3KDS was developed. The results show that SPT kinetics in response …