PMID- 11799124
OWN - NLM
STAT- MEDLINE
DCOM- 20020513
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 277
IP  - 14
DP  - 2002 Apr 5
TI  - Hepatocyte growth factor/scatter factor binds to small heparin-derived 
      oligosaccharides and stimulates the proliferation of human HaCaT keratinocytes.
PG  - 12456-62
AB  - Hepatocyte growth factor/scatter factor (HGF/SF) acts via a dual receptor system 
      consisting of the MET tyrosine kinase receptor and heparan sulfate or dermatan 
      sulfate proteoglycans. In optical biosensor binding assays, competition by 
      oligosaccharides for binding of HGF/SF to immobilized heparin showed that 
      disaccharides failed to compete, whereas tetrasaccharides inhibited HGF/SF 
      binding (IC(50) 8 microg/ml). The inhibitory potency of the oligosaccharides 
      increased as their length increased by successive disaccharide units, to reach a 
      maximum (IC(50) 1 microg/ml) at degree of polymerization (dp) 10. In binding 
      assays, HGF/SF was found to bind directly to oligosaccharides as small as dp 4, 
      and the binding parameters were similar for oligosaccharides of dp 4-14 (k(a) 
      2.2-45.3 x 10(6) m(-1) s(-1), k(d) 0.033-0.039 s(-1), and K(d) 9-16 nm). In human 
      keratinocytes, HGF/SF stimulated DNA synthesis, and this was dependent on a 
      sustained phosphorylation of p42/44(MAPK). In chlorate-treated and hence sulfated 
      glycosaminoglycan-deficient HaCaT cells, the stimulation of DNA synthesis by 
      HGF/SF was almost abolished. Heparin-derived oligosaccharides from dp 2 to dp 24 
      were added together with HGF/SF to chlorate-treated cells to determine the 
      minimum size of oligosaccharides able to restore HGF/SF activity. At restricted 
      concentrations of oligosaccharides (4 ng/ml), HGF/SF required decasaccharides, 
      whereas at higher concentrations (100 ng/ml) even tetrasaccharides were able to 
      partly restore DNA synthesis. The results suggest that HGF/SF binds to a 
      tetrasaccharide and that although this is sufficient to enable the stimulation of 
      DNA synthesis, longer oligosaccharides are more efficient, perhaps by virtue of 
      their ability to bind more easily other molecules.
FAU - Delehedde, Maryse
AU  - Delehedde M
AD  - School of Biological Sciences, Life Science Building, University of Liverpool, 
      Crown Street, Liverpool L69 7ZB, United Kingdom.
FAU - Lyon, Malcolm
AU  - Lyon M
FAU - Vidyasagar, Rishma
AU  - Vidyasagar R
FAU - McDonnell, Timothy J
AU  - McDonnell TJ
FAU - Fernig, David G
AU  - Fernig DG
LA  - eng
GR  - P01 CA68233/CA/NCI 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.
DEP - 20020117
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Chlorates)
RN  - 0 (Glycosaminoglycans)
RN  - 0 (Oligosaccharides)
RN  - 67256-21-7 (Hepatocyte Growth Factor)
RN  - 9005-49-6 (Heparin)
RN  - 9007-49-2 (DNA)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 1)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinase 3)
RN  - EC 2.7.11.24 (Mitogen-Activated Protein Kinases)
SB  - IM
MH  - Binding, Competitive
MH  - Biosensing Techniques
MH  - Biotinylation
MH  - Cell Division
MH  - Chlorates/pharmacology
MH  - DNA/biosynthesis
MH  - Dimerization
MH  - Dose-Response Relationship, Drug
MH  - Glycosaminoglycans/chemistry
MH  - Heparin/*metabolism
MH  - Hepatocyte Growth Factor/*chemistry/*metabolism
MH  - Humans
MH  - Inhibitory Concentration 50
MH  - Keratinocytes/*metabolism
MH  - Kinetics
MH  - Mitogen-Activated Protein Kinase 1/metabolism
MH  - Mitogen-Activated Protein Kinase 3
MH  - Mitogen-Activated Protein Kinases/metabolism
MH  - Oligosaccharides/*chemistry
MH  - Phosphorylation
MH  - Protein Binding
MH  - Protein Structure, Tertiary
MH  - Time Factors
EDAT- 2002/01/19 10:00
MHDA- 2002/05/15 10:01
CRDT- 2002/01/19 10:00
PHST- 2002/01/19 10:00 [pubmed]
PHST- 2002/05/15 10:01 [medline]
PHST- 2002/01/19 10:00 [entrez]
AID - S0021-9258(18)52078-0 [pii]
AID - 10.1074/jbc.M111345200 [doi]
PST - ppublish
SO  - J Biol Chem. 2002 Apr 5;277(14):12456-62. doi: 10.1074/jbc.M111345200. Epub 2002 
      Jan 17.