Unveiling ceramide synthase 2 (CerS2): From characteristics and isolation to enzyme activity assays

Abstract

Ceramide synthase 2 (CerS2) is an essential enzyme in the metabolic pathway of sphingolipids, a class of membrane lipids that act crucial roles in various cellular functions [1]. Among the six mammalian CerS enzymes, CerS2 stands out for its omnipresence and abundant expression in various mammalian tissues and organs. CerS2 mRNA and protein expression levels are particularly prominent in the kidney, liver, lung, intestine, and other essential tissues [1–3]. This widespread distribution underscores CerS2's significance as a critical contributor to basal cellular sphingolipid metabolism. The enzyme's substrate specificity for specific acyl-CoAs, such as C20:0, C22:0, C24:0, C24:1, and C26:0, further accentuates its role in generating distinct ceramide species. CerS2's presence has been identified in the endoplasmic reticulum (ER), where it catalyzes the synthesis of ceramides by combining fatty-acyl chains with dihydrosphingosine. Analysis of CerS2 expression and activity involves a combination of techniques, when the real-time PCR measures mRNA levels, thus providing insights into the transcriptional regulation of CerS2. Western blotting is used to determine the protein abundance of CerS2 on its overall protein levels. In this review, we offer a thorough analysis of CerS2, encompassing its distinctive characteristics, biological relevance, isolation methods, quantification at both the mRNA and protein levels, as well as an assessment of its enzymatic activity via appropriate assays.