PMID- 10514476
OWN - NLM
STAT- MEDLINE
DCOM- 19991119
LR  - 20210209
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 42
DP  - 1999 Oct 15
TI  - Interaction of macrophage-stimulating protein with its receptor. Residues 
      critical for beta chain binding and evidence for independent alpha chain binding.
PG  - 29937-43
AB  - Macrophage-stimulating protein (MSP) and hepatocyte growth factor/scatter factor 
      (HGF/SF) are plasminogen-related growth and motility factors that interact with 
      cell-surface protein tyrosine kinase receptors. Each one is a heterodimeric 
      protein comprising a disulfide-linked alpha chain and a serine protease-like beta 
      chain. Despite structural similarities between MSP and HGF, the primary receptor 
      binding site is located on the alpha chain of HGF/SF but on the beta chain of 
      MSP. To obtain insight into the structural basis for MSP beta chain binding, beta 
      chain structure was modeled from coordinates of an existing model of the HGF beta 
      chain. The model revealed that the region corresponding to the S1 specificity 
      pocket in trypsin is filled by the Asn(682)/Glu(648) interacting pair, leaving a 
      shallow cavity for possible beta chain interaction with the receptor. Mutants in 
      this region were created, and their binding characteristics were determined. A 
      double mutation of Asn(682)/Glu(648) caused diminished binding of the beta chain 
      to the MSP receptor, and a single mutation of neighboring Arg(683) completely 
      abolished binding. Thus, this region of the molecule is critical for binding. We 
      also found that at equimolar concentrations of free alpha and beta chains, alpha 
      chain binding to receptor was detectable, at levels considerably lower than beta 
      chain binding. The EC(50) values determined by quantitative enzyme-linked 
      immunosorbent assay are 0.25 and 16.9 nM for beta and alpha chain, respectively. 
      The data suggest that MSP has two independent binding sites with high and low 
      affinities located in beta and alpha chain, respectively, and that the two sites 
      together mediate receptor dimerization and subsequent activation.
FAU - Danilkovitch, A
AU  - Danilkovitch A
AD  - Laboratory of Immunobiology, NCI-Frederick Cancer Research and Development 
      Center, ABL-Basic Research Program, Frederick, Maryland 21702, USA. 
      danilkovitch@mail.ncifcrf.gov
FAU - Miller, M
AU  - Miller M
FAU - Leonard, E J
AU  - Leonard EJ
LA  - eng
PT  - Journal Article
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 (DNA, Complementary)
RN  - 0 (Growth Substances)
RN  - 0 (Proto-Oncogene Proteins)
RN  - 0 (macrophage stimulating protein)
RN  - 67256-21-7 (Hepatocyte Growth Factor)
RN  - EC 2.7.10.1 (Proto-Oncogene Proteins c-met)
RN  - EC 3.4.21.- (Serine Endopeptidases)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - CHO Cells
MH  - Cell Line
MH  - Cricetinae
MH  - DNA, Complementary
MH  - Dogs
MH  - Growth Substances/chemistry/genetics/*metabolism
MH  - *Hepatocyte Growth Factor
MH  - Molecular Sequence Data
MH  - Protein Binding
MH  - *Proto-Oncogene Proteins
MH  - Proto-Oncogene Proteins c-met/*metabolism
MH  - Sequence Homology, Amino Acid
MH  - Serine Endopeptidases/metabolism
EDAT- 1999/10/09 00:00
MHDA- 1999/10/09 00:01
CRDT- 1999/10/09 00:00
PHST- 1999/10/09 00:00 [pubmed]
PHST- 1999/10/09 00:01 [medline]
PHST- 1999/10/09 00:00 [entrez]
AID - S0021-9258(19)51849-X [pii]
AID - 10.1074/jbc.274.42.29937 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Oct 15;274(42):29937-43. doi: 10.1074/jbc.274.42.29937.