PMID- 9443912
OWN - NLM
STAT- MEDLINE
DCOM- 19980409
LR  - 20211203
IS  - 0960-9822 (Print)
IS  - 0960-9822 (Linking)
VI  - 8
IP  - 3
DP  - 1998 Jan 29
TI  - Engineered mutants of HGF/SF with reduced binding to heparan sulphate 
      proteoglycans, decreased clearance and enhanced activity in vivo.
PG  - 125-34
AB  - BACKGROUND: Although a number of growth factors bind cell-surface heparan 
      sulphate proteoglycans (HSPGs), the role of this interaction is unclear except 
      for fibroblast growth factor which requires HSPG binding for signalling. 
      Hepatocyte growth factor/scatter factor (HGF/SF) plays important roles in 
      mammalian development and tissue regeneration and acts on target cells through a 
      specific receptor tyrosine kinase encoded by the c-met proto-oncogene. This 
      factor also binds HSPGs with high affinity, but conflicting data have been 
      reported on the role of HSPG binding in HGF/SF signalling. RESULTS: To map the 
      binding sites for HSPG and the Met receptor in HGF/SF, we have engineered a 
      number of HGF/SF mutants in which several clusters of solvent-accessible residues 
      in the hairpin structure of the amino-terminal domain or in kringle 2 have been 
      replaced. Two of the mutants (HP1 and HP2) showed greatly decreased (more than 
      50-fold) affinity for heparin and HSPGs but retained full mitogenic and motogenic 
      activities on target cells in culture. Furthermore, when compared with wild-type 
      HGF/SF, the HP1 mutant exhibited a delayed clearance from the blood, higher 
      tissue levels and a higher induction of DNA synthesis in normal, adult murine 
      liver. CONCLUSIONS: These results establish the following: the binding sites in 
      HGF/SF for Met and for HSPGs can be dissociated by protein engineering; 
      high-affinity binding of HGF/SF to HSPGs is not essential for signalling; one 
      role of HSPG binding in the HGF/SF system appears to be sequestration and 
      degradation of the growth factor; and HGF/SF mutants with decreased affinity for 
      HSPGs exhibit enhanced activity in vivo.
FAU - Hartmann, G
AU  - Hartmann G
AD  - Imperial Cancer Research Fund Laboratory, MRC Centre, Hills Road, Cambridge, CB2 
      2QH, UK.
FAU - Prospero, T
AU  - Prospero T
FAU - Brinkmann, V
AU  - Brinkmann V
FAU - Ozcelik, C
AU  - Ozcelik C
FAU - Winter, G
AU  - Winter G
FAU - Hepple, J
AU  - Hepple J
FAU - Batley, S
AU  - Batley S
FAU - Bladt, F
AU  - Bladt F
FAU - Sachs, M
AU  - Sachs M
FAU - Birchmeier, C
AU  - Birchmeier C
FAU - Birchmeier, W
AU  - Birchmeier W
FAU - Gherardi, E
AU  - Gherardi E
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Curr Biol
JT  - Current biology : CB
JID - 9107782
RN  - 0 (Heparan Sulfate Proteoglycans)
RN  - 0 (MAS1 protein, human)
RN  - 0 (Peptide Fragments)
RN  - 0 (Proto-Oncogene Mas)
RN  - 0 (Recombinant Fusion Proteins)
RN  - 67256-21-7 (Hepatocyte Growth Factor)
RN  - 9005-49-6 (Heparin)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-met)
SB  - IM
EIN - Curr Biol 1998 Oct 8;8(20):R739. Ozcelik O [corrected to Ozcelik C]
MH  - Animals
MH  - Binding Sites
MH  - Cell Line
MH  - DNA Replication/drug effects
MH  - Dogs
MH  - Female
MH  - Heparan Sulfate Proteoglycans/*metabolism
MH  - Heparin/metabolism
MH  - Hepatocyte Growth 
      Factor/chemistry/*genetics/metabolism/pharmacokinetics/pharmacology
MH  - Humans
MH  - Kringles/genetics
MH  - Liver/metabolism
MH  - Metabolic Clearance Rate
MH  - Mink
MH  - Models, Molecular
MH  - Mutagenesis, Site-Directed
MH  - Peptide Fragments/metabolism
MH  - Protein Binding
MH  - Protein Conformation
MH  - Proto-Oncogene Mas
MH  - Proto-Oncogene Proteins c-met/*metabolism
MH  - Rats
MH  - Recombinant Fusion Proteins/chemistry/metabolism/pharmacokinetics/pharmacology
MH  - Tissue Distribution
EDAT- 1998/04/16 00:00
MHDA- 1998/04/16 00:01
CRDT- 1998/04/16 00:00
PHST- 1998/04/16 00:00 [pubmed]
PHST- 1998/04/16 00:01 [medline]
PHST- 1998/04/16 00:00 [entrez]
AID - S0960-9822(98)70059-4 [pii]
AID - 10.1016/s0960-9822(98)70059-4 [doi]
PST - ppublish
SO  - Curr Biol. 1998 Jan 29;8(3):125-34. doi: 10.1016/s0960-9822(98)70059-4.