PMID- 10455156
OWN - NLM
STAT- MEDLINE
DCOM- 19990930
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 35
DP  - 1999 Aug 27
TI  - A novel glycosulfopeptide binds to P-selectin and inhibits leukocyte adhesion to 
      P-selectin.
PG  - 24838-48
AB  - P-selectin glycoprotein ligand-1 (PSGL-1) is a dimeric membrane mucin on
      leukocytes that binds selectins. The molecular features of PSGL-1 that determine 
      this high affinity binding are unclear. Here we demonstrate the in vitro
      synthesis of a novel glycosulfopeptide (GSP-6) modeled after the extreme N
      terminus of PSGL-1, which has been predicted to be important for P-selectin
      binding. GSP-6 contains three tyrosine sulfate (TyrSO(3)) residues and a
      monosialylated, core 2-based O-glycan with a sialyl Lewis x (C2-O-sLe(x)) motif
      at a specific Thr residue. GSP-6 binds tightly to immobilized P-selectin, whereas
      glycopeptides lacking either TyrSO(3) or C2-O-sLe(x) do not detectably bind.
      Remarkably, an isomeric glycosulfopeptide to GSP-6, termed GSP-6', which contains
      sLe(x) on an extended core 1-based O-glycan, does not bind immobilized
      P-selectin. Equilibrium gel filtration analysis revealed that GSP-6 binds to
      soluble P-selectin with a K(d) of approximately 350 nM. GSP-6 (<5 microM)
      substantially inhibits neutrophil adhesion to P-selectin in vitro, whereas free
      sLe(x) (5 mM) only slightly inhibits adhesion. In contrast to the inherent
      heterogeneity of post-translational modifications of recombinant proteins,
      glycosulfopeptides permit the placement of sulfate groups and glycans of precise 
      structure at defined positions on a polypeptide. This approach should expedite
      the probing of structure-function relationships in sulfated and glycosylated
      proteins, and may facilitate development of novel drugs to treat inflammatory
      diseases involving P-selectin-mediated leukocyte adhesion.
FAU - Leppanen, A
AU  - Leppanen A
AD  - Department of Biochemistry and Molecular Biology, Oklahoma City, Oklahoma 73104, 
      USA.
FAU - Mehta, P
AU  - Mehta P
FAU - Ouyang, Y B
AU  - Ouyang YB
FAU - Ju, T
AU  - Ju T
FAU - Helin, J
AU  - Helin J
FAU - Moore, K L
AU  - Moore KL
FAU - van Die, I
AU  - van Die I
FAU - Canfield, W M
AU  - Canfield WM
FAU - McEver, R P
AU  - McEver RP
FAU - Cummings, R D
AU  - Cummings RD
LA  - eng
GR  - P01 HL54804/HL/NHLBI NIH HHS/United States
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, P.H.S.
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Carrier Proteins)
RN  - 0 (GSP-6 glycosulfopeptide)
RN  - 0 (Glycoproteins)
RN  - 0 (Lewis X Antigen)
RN  - 0 (Membrane Glycoproteins)
RN  - 0 (P-Selectin)
RN  - 0 (P-selectin ligand protein)
RN  - 0 (Peptides)
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Amino Acid Sequence
MH  - Carrier Proteins/*chemical synthesis/pharmacology
MH  - Cell Adhesion/*drug effects
MH  - Chromatography, Affinity
MH  - Chromatography, High Pressure Liquid
MH  - Dimerization
MH  - *Glycoproteins
MH  - Humans
MH  - Lewis X Antigen/chemistry
MH  - Mass Spectrometry
MH  - Membrane Glycoproteins/*chemical synthesis/chemistry/*pharmacology
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Neutrophils/*metabolism
MH  - P-Selectin/*metabolism
MH  - *Peptides
MH  - Polysaccharides/chemistry
MH  - Protein Binding
EDAT- 1999/08/24 00:00
MHDA- 1999/08/24 00:01
CRDT- 1999/08/24 00:00
PHST- 1999/08/24 00:00 [pubmed]
PHST- 1999/08/24 00:01 [medline]
PHST- 1999/08/24 00:00 [entrez]
AID - 10.1074/jbc.274.35.24838 [doi]
AID - S0021-9258(19)55422-9 [pii]
PST - ppublish
SO  - J Biol Chem. 1999 Aug 27;274(35):24838-48. doi: 10.1074/jbc.274.35.24838.