PMID- 10200296
OWN - NLM
STAT- MEDLINE
DCOM- 19990517
LR  - 20190501
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 96
IP  - 8
DP  - 1999 Apr 13
TI  - Reconstitution of functional L-selectin ligands on a cultured human endothelial
      cell line by cotransfection of alpha1-->3 fucosyltransferase VII and newly cloned
      GlcNAcbeta:6-sulfotransferase cDNA.
PG  - 4530-5
AB  - Recently, we proposed sialyl 6-sulfo Lewis X as a major carbohydrate-capping
      group of the L-selectin ligands on high endothelial venules in human lymph nodes.
      In this study we succeeded in reconstituting functional L-selectin ligands on a
      cultured human endothelial cell line, ECV304, by transfecting the
      alpha1-->3fucosyltranseferase VII (Fuc-T VII) and newly cloned
      GlcNAcbeta:6-sulfotransferase (6-Sul-T) cDNAs. The ECV304 cells transfected with 
      Fuc-T VII cDNA expressed conventional sialyl Lewis X detected with specific
      antibodies including 2H5, whereas the cells transfected with 6-Sul-T cDNA
      expressed sialyl 6-sulfo lactosamine as well as MECA-79-defined carbohydrate
      determinants, but these singly transfected cells failed to express sialyl 6-sulfo
      Lewis X, as detected with the antisialyl 6-sulfo Lewis X mAb G152. Sialyl 6-sulfo
      Lewis X appeared only on the cells that were cotransfected with both 6-Sul-T and 
      Fuc-T VII cDNAs. Significant adhesion of L-selectin-expressing cells was seen
      only to the doubly transfected ECV304 cells and was inhibited by G152. No
      adhesion was observed to the cells transfected either with 6-Sul-T or with Fuc-T 
      VII cDNA alone. The mRNAs of the two enzymes were expressed or were inducible
      upon interleukin 1 stimulation in human endothelial cells. These results indicate
      that a set of carbohydrate determinants synthesized by the concerted action of
      the two enzymes, as typically represented by the sialyl 6-sulfo Lewis X-capping
      group, serves as an essential component of the ligand for L-selectin and that the
      reagents 2H5 and MECA-79, utilized in earlier studies to detect L-selectin ligand
      on high endothelial venules, recognize two different aspects of the same set of
      synthetic products.
FAU - Kimura, N
AU  - Kimura N
AD  - Program of Experimental Pathology, Aichi Cancer Center, Nagoya 464-8681, Japan.
FAU - Mitsuoka, C
AU  - Mitsuoka C
FAU - Kanamori, A
AU  - Kanamori A
FAU - Hiraiwa, N
AU  - Hiraiwa N
FAU - Uchimura, K
AU  - Uchimura K
FAU - Muramatsu, T
AU  - Muramatsu T
FAU - Tamatani, T
AU  - Tamatani T
FAU - Kansas, G S
AU  - Kansas GS
FAU - Kannagi, R
AU  - Kannagi R
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0
      (5-acetylneuraminyl-(2-3)-galactosyl-(1-4)-(fucopyranosyl-(1-3))-N-acetylglucosam
      ine)
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (DNA, Complementary)
RN  - 0 (Oligosaccharides)
RN  - 126880-86-2 (L-Selectin)
RN  - EC 2.4.1.- (Fucosyltransferases)
RN  - EC 2.4.1.152 (galactoside 3-fucosyltransferase)
RN  - EC 2.8.2.- (Sulfotransferases)
RN  - EC 2.8.2.- (carbohydrate sulfotransferases)
SB  - IM
MH  - Antibodies, Monoclonal
MH  - Carbohydrate Sequence
MH  - Cell Line
MH  - Cloning, Molecular
MH  - DNA, Complementary
MH  - Endothelium, Vascular/cytology/*physiology
MH  - Fucosyltransferases/genetics/*metabolism
MH  - Genetic Variation
MH  - Humans
MH  - L-Selectin/*physiology
MH  - Lymph Nodes/immunology
MH  - Molecular Sequence Data
MH  - Oligosaccharides/*biosynthesis/chemistry/genetics
MH  - Sulfotransferases/genetics/*metabolism
MH  - Transfection
MH  - Umbilical Veins
PMC - PMC16366
EDAT- 1999/04/14 00:00
MHDA- 1999/04/14 00:01
CRDT- 1999/04/14 00:00
PHST- 1999/04/14 00:00 [pubmed]
PHST- 1999/04/14 00:01 [medline]
PHST- 1999/04/14 00:00 [entrez]
AID - 10.1073/pnas.96.8.4530 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 1999 Apr 13;96(8):4530-5. doi: 10.1073/pnas.96.8.4530.