Hofmeister, Robert and Boettcher, Alfred and Schmitz, Gerd (1998) Preparation of Golgi subfractions with free-solution isotachophoresis: Analysis of sphingomyelin synthesis in Golgi subfractions from rat liver. ELECTROPHORESIS, 19 (7). pp. 1185-1194. ISSN 0173-0835,
Full text not available from this repository. (Request a copy)Abstract
A new displacement electrophoresis technique, termed free-solution isotachophoresis (FS-ITP) was used for the analysis of sphingolipid metabolism in Golgi subfractions. The discontinuous electrolyte system enables tissue-derived membrane vesicles to be separated and purified due to their polarity patterns in a mobility gradient. In this study total Golgi apparatus obtained from rat liver by discontinuous density gradient centrifugation was subfractionated by preparative FS-ITP, yielding enzymatically active cis-, medial-, and trans-Golgi subfractions. These membrane vesicles were assayed by the following established enzyme marker activities: NADH cytochrome c reductase (cis-Golgi), NADP phosphatase (medial-Golgi), and thiamine pyrophosphatase (trans-Golgi). The activity of phosphatidylcholine :ceramide phosphocholine transferase, a sphingomyelin synthesizing enzyme, is attributed to the cis- and medial-Golgi-derived subfractions. Analysis of Golgi lipids revealed a decline in membranous ceramide along the cis- to trans-Golgi polarity axis. Furthermore, significant amounts of newly synthesized sphingomyelin and diacylglycerol are transferred from the medial/cis- to the trans-Golgi compartment. The FS-ITP system is well suited for micropreparative experimental applications, as demonstrated by studies on phosphatidylcholine :ceramide phosphocholine transferase activity in Golgi membrane vesicles of rat liver obtained by FS-ITP.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | PROTEIN-KINASE-C; SUBCELLULAR-LOCALIZATION; ENDOPLASMIC-RETICULUM; PLASMA-MEMBRANES; APPARATUS; PHOSPHATIDYLCHOLINE; BIOSYNTHESIS; CERAMIDE; LIPOPROTEINS; CHOLESTEROL; preparative isotachophoresis; Golgi apparatus; sphingomyelin synthesis; phosphatidylcholine, ceramide phosphocholine transferase |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Klinische Chemie und Laboratoriumsmedizin |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 09 May 2023 09:09 |
| Last Modified: | 09 May 2023 09:09 |
| URI: | https://pred.uni-regensburg.de/id/eprint/49752 |
Actions (login required)
 |
View Item |
