Linked Life Data

7680530

Source:http://linkedlifedata.com/resource/pubmed/id/7680530

Statements in which the resource exists as a subject.
PredicateObject
rdf:type
lifeskim:mentions
pubmed:issue
2 Pt 1
pubmed:dateCreated
1993-4-2
pubmed:abstractText
To distinguish cystic fibrosis transmembrane conductance regulator (CFTR) at the surface of epithelial cells from that present in intracellular membranes, intact T84 cells were treated with periodate and biotin-LC-hydrazide to derivatize exposed glycoconjugates. Cell lysates were then passed over a monomeric avidin column, which allows reversible avidin-biotin binding. After washing, biotinylated molecules were eluted with 2 mM biotin. CFTR was then immunoprecipitated with a mouse monoclonal antibody from both the unbound and biotin eluent fractions, radioactively labeled by in vitro phosphorylation, and analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Nonimmune mouse immunoglobulin G failed to precipitate any CFTR, and CFTR was detectable only in the wash fractions when cells were periodate treated but not labeled with biotin hydrazide. In biotinylated cells, CFTR levels were approximately equal in the unbound fraction and the biotin eluent. The proportion of biotinylated CFTR did not significantly increase when cells were labeled after treatment with 10 microM forskolin. These data demonstrate that in T84 cells CFTR is constitutively expressed on the cell surface and that activation of CFTR does not primarily depend on the cAMP-dependent trafficking of CFTR to the plasma membrane. The large unbiotinylated pool of maturely glycosylated CFTR suggests that CFTR resides in an intracellular compartment as well as being present at the cell surface.
pubmed:grant
pubmed:language
eng
pubmed:journal
pubmed:citationSubset
IM
pubmed:chemical
pubmed:status
MEDLINE
pubmed:month
Feb
pubmed:issn
0002-9513
pubmed:author
pubmed:issnType
Print
pubmed:volume
264
pubmed:owner
NLM
pubmed:authorsComplete
Y
pubmed:pagination
C491-8
pubmed:dateRevised
2007-11-14
pubmed:meshHeading
pubmed:year
1993
pubmed:articleTitle
Cell surface labeling of CFTR in T84 cells.
pubmed:affiliation
Gregory Fleming James Cystic Fibrosis Research Center, University of Alabama, Birmingham 35294.
pubmed:publicationType
Journal Article, Research Support, U.S. Gov't, P.H.S., Research Support, Non-U.S. Gov't