Density-labelling of cell wall polysaccharides in cultured rose cells: comparison of incorporation of H-2 and C-13 from exogenous glucose

J E Thompson, S C Fry

Research output: Contribution to journalArticlepeer-review

Abstract / Description of output

Labelling with stable isotopes has under-exploited potential for studies of polysaccharide endotransglycosylation in vivo. Ideally, the labelled polysaccharides should have the highest possible buoyant density. Although [C-13(6)]glucose has previously been used as a precursor, it was unclear whether H-2 would be efficiently incorporated from [H-2]glucose or lost as D2O. Rose (Rosa sp.) cell-suspension cultures efficiently incorporated C-13 from D-[C-13(6),H-2(7)]glucose into wall polysaccharides with negligible dilution from atmospheric (CO2)-C-12. Also, similar to 70% of the H-2 atoms in D-[C-13(6),H-2(7)]glucose were retained during polysaccharide biosynthesis. This shows that relatively few cycles of intermediary metabolism leading to the release of D2O occurred before sugar residues were incorporated into wall polysaccharides. In agreement with these observations, isopycnic centrifugation in caesium trifluoroacetate gradients showed that the hydrated buoyant density of xyloglucan synthesised by rose cells growing on [C-13(6),H-2(7)]glucose and [C-13(6)]glucose was 3.7 and 2.6% higher, respectively, than in isotopically non-labelled cultures. Thus, [C-13,H-2]glucosefeeding enabled a 42% better resolution of 'heavy' from 'light' xyloglucan than [C-13]glucose-feeding.

Original languageEnglish
Pages (from-to)175-182
Number of pages8
JournalCarbohydrate Research
Volume332
Issue number2
DOIs
Publication statusPublished - 18 May 2001

Keywords / Materials (for Non-textual outputs)

  • Xyloglucan
  • Biosynthesis
  • Isopycnic centrifugation
  • Caesium trifluoroacetate (CsTFA™)
  • Transglycosylation
  • Density-labelling

Fingerprint

Dive into the research topics of 'Density-labelling of cell wall polysaccharides in cultured rose cells: comparison of incorporation of H-2 and C-13 from exogenous glucose'. Together they form a unique fingerprint.

Cite this