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PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
14
pubmed:dateCreated
1992-6-9
pubmed:abstractText
FP21 is a 21-kDa fucoprotein which fractionates with the cytosol after high-speed centrifugation of gently lysed Dictyostelium cells. Less than 0.7% of FP21 is associated with vesicles. In proliferating cells, 4 x 10(5) fucosyl moieties/cell are associated with FP21 as anionic, possibly O-linked oligosaccharides equal in size to 4.8 glucose units. FP21 is underfucosylated in a mutant strain (HL250) that depends on extracellular fucose for synthesis of GDP-fucose. To determine the cellular site of FP21 fucosylation, cytosolic and vesicular preparations from strain HL250 were compared for their ability to transfer fucose from GDP-fucose to FP21. Cytosolic preparations fucosylate endogenous FP21 in a time-, concentration-, and divalent cation-dependent fashion, with a Km for GDP-fucose of 1.4 microM. Activity in normal cell cytosol is dependent on exogenous mutant FP21, demonstrating that FP21 is normally fully fucosylated. Both mutant and normal cytosols are also able to alpha-fucosylate a type 1 glycolipid substrate (8-methoxycarbonyloctyl-Gal beta 1-3 beta GlcNAc), but not related substrates, with Km values for the type 1 glycolipid of 0.99 mM and for GDP-fucose of 1.6 microM. Competitive inhibition between FP21 and the type 1 glycolipid shows that the same enzyme fucosylates both substrates. Intact and permeabilized vesicle preparations from wild-type cells are unable to fucosylate FP21 or the type 1 glycolipid by a divalent cation-dependent mechanism, and thus are devoid of FP21-fucosyltransferase. Since control experiments showed that vesicle leakage is minimal during cytosol preparation, these results indicate that FP21 is synthesized and fucosylated in the cytosolic compartment, by an unusual soluble fucosyltransferase.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
May
pubmed:issn
0021-9258
pubmed:author
pubmed:issnType
Print
pubmed:day
15
pubmed:volume
267
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
9595-605
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1992
pubmed:articleTitle
Characterization of a cytosolic fucosylation pathway in Dictyostelium.
pubmed:affiliation
Department of Anatomy and Cell Biology, College of Medicine, University of Florida, Gainesville.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S.