PMID- 15128268
OWN - NLM
STAT- MEDLINE
DCOM- 20041210
LR  - 20181113
IS  - 1470-8728 (Electronic)
IS  - 0264-6021 (Print)
IS  - 0264-6021 (Linking)
VI  - 381
IP  - Pt 3
DP  - 2004 Aug 1
TI  - Cellular effects of deoxynojirimycin analogues: uptake, retention and inhibition 
      of glycosphingolipid biosynthesis.
PG  - 861-6
AB  - Deoxynojirimycin (DNJ) analogues are inhibitors of ceramide glucosyltransferase 
      (CGT), which catalyses the first step in the glucosphingolipid (GSL) biosynthetic 
      pathway. We have synthesized a series of DNJ analogues to study the contribution 
      of N-alk(en)yl side chains (C4, C9 or C18) to the behaviour of these analogues in 
      cultured HL60 cells. When cells were treated for 16 h at non-cytotoxic 
      concentrations of inhibitor, a 40-50% decrease in GSL levels was measured by HPLC 
      analysis of GSL-derived oligosaccharides following ceramide glycanase digestion 
      of GSL and 2-aminobenzamide labelling of the released oligosaccharides. Using a 
      novel technique for short-term [14C]galactose labelling of cellular GSL, we used 
      compound inhibition of GSL biosynthesis as a marker for compound uptake into 
      cells. Surprisingly, the uptake of all three of the DNJ analogues was extremely 
      rapid and was not dependent upon the length of the N-alk(en)yl moiety. Compound 
      uptake occurred in less than 1 min, as shown by the complete inhibition of GSL 
      labelling in cells treated with all the DNJ analogues. Greatly increased cellular 
      retention of N-cis-13-octadecenyl-DNJ was observed relative to the shorter-chain 
      compounds, N-butyl-DNJ and N-nonyl-DNJ, as indicated by complete inhibition of 
      CGT 24 h after removal of inhibitor from the culture medium. The present study 
      further characterizes the properties of N-alk(en)ylated DNJs, and demonstrates 
      that increasing the length of the side chain is a simple way of improving imino 
      sugar retention and therefore inhibitory efficacy for CGT in cultured cells.
FAU - Mellor, Howard R
AU  - Mellor HR
AD  - Glycobiology Institute, Department of Biochemistry, University of Oxford, South 
      Parks Road, Oxford OX1 3QU, UK.
FAU - Neville, David C A
AU  - Neville DC
FAU - Harvey, David J
AU  - Harvey DJ
FAU - Platt, Frances M
AU  - Platt FM
FAU - Dwek, Raymond A
AU  - Dwek RA
FAU - Butters, Terry D
AU  - Butters TD
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Biochem J
JT  - The Biochemical journal
JID - 2984726R
RN  - 0 (Carbon Radioisotopes)
RN  - 0 (Enzyme Inhibitors)
RN  - 0 (Glycosphingolipids)
RN  - 19130-96-2 (1-Deoxynojirimycin)
SB  - IM
MH  - 1-Deoxynojirimycin/*analogs & derivatives/metabolism/*pharmacology
MH  - Carbon Radioisotopes/metabolism
MH  - Cell Line, Tumor
MH  - Chromatography, High Pressure Liquid
MH  - Enzyme Inhibitors/metabolism/pharmacology
MH  - Glycosphingolipids/*antagonists & inhibitors/*biosynthesis/metabolism
MH  - HL-60 Cells/chemistry/*drug effects/*metabolism
MH  - Humans
MH  - Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
MH  - Staining and Labeling/methods
PMC - PMC1133897
EDAT- 2004/05/07 05:00
MHDA- 2004/12/16 09:00
PMCR- 2005/02/01
CRDT- 2004/05/07 05:00
PHST- 2004/05/06 00:00 [accepted]
PHST- 2004/04/21 00:00 [revised]
PHST- 2003/11/27 00:00 [received]
PHST- 2004/05/07 05:00 [pubmed]
PHST- 2004/12/16 09:00 [medline]
PHST- 2004/05/07 05:00 [entrez]
PHST- 2005/02/01 00:00 [pmc-release]
AID - BJ20031822 [pii]
AID - bj3810861 [pii]
AID - 10.1042/BJ20031822 [doi]
PST - ppublish
SO  - Biochem J. 2004 Aug 1;381(Pt 3):861-6. doi: 10.1042/BJ20031822.