PMID- 8631822
OWN - NLM
STAT- MEDLINE
DCOM- 19960703
LR  - 20220310
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 271
IP  - 13
DP  - 1996 Mar 29
TI  - The carboxyl-terminal domain (111-165) of vascular endothelial growth factor is 
      critical for its mitogenic potency.
PG  - 7788-95
AB  - Vascular endothelial growth factor (VEGF) is a potent and specific mitogen for 
      endothelial cells. VEGF is synthesized and secreted by many differentiated cells 
      in response to a variety of stimuli including hypoxia. VEGF is expressed in a 
      variety of tissues as multiple homodimeric forms (121, 165, 189, and 206 amino 
      acids/monomer) resulting from alternative RNA splicing. VEGF121 is a soluble 
      mitogen that does not bind heparin; the longer forms of VEGF bind heparin with 
      progressively higher affinity. The higher molecular weight forms of VEGF can be 
      cleaved by plasmin to release a diffusible form(s) of VEGF. We characterized the 
      proteolysis of VEGF by plasmin and other proteases. Thrombin, elastase, and 
      collagenase did not cleave VEGF, whereas trypsin generated a series of smaller 
      fragments. The isolated plasmin fragments of VEGF were compared with respect to 
      heparin binding, interaction with soluble VEGF receptors, and ability to promote 
      endothelial cell mitogenesis. Plasmin yields two fragments of VEGF as indicated 
      by reverse phase high performance liquid chromatography and SDS-polyacrylamide 
      gel electrophoresis: an amino-terminal homodimeric protein containing receptor 
      binding determinants and a carboxyl-terminal polypeptide which bound heparin. 
      Amino-terminal sequencing of the carboxyl-terminal peptide identified the plasmin 
      cleavage site as Arg110-Ala111. A heterodimeric form of VEGF165/110, was isolated 
      from partial plasmin digests of VEGF165. The carboxyl-terminal polypeptide 
      (111-165) displayed no affinity for soluble kinase domain region (KDR) or 
      Fms-like tyrosine kinase (FLT-1) receptors. The various isoforms of VEGF (165, 
      165/110, and 121) bound soluble kinase domain region receptor with similar 
      affinity (approximately 30 pM). In contrast, soluble FLT-1 receptor 
      differentiated VEGF isoforms (165, 165/110, 110, and 121) with apparent 
      affinities of 10, 30, 120, and 200 pM, respectively. Endothelial cell mitogenic 
      potencies of VEGF110 and VEGF121 were decreased more than 100-fold compared to 
      that of VEGF165. The present findings indicate that removal of the 
      carboxyl-terminal domain, whether it is due to alternative splicing of mRNA or to 
      proteolysis, is associated with a significant loss in bioactivity.
FAU - Keyt, B A
AU  - Keyt BA
AD  - Department of Cardiovascular Research, Genentech, Inc., South San Francisco, 
      California 94080, USA.
FAU - Berleau, L T
AU  - Berleau LT
FAU - Nguyen, H V
AU  - Nguyen HV
FAU - Chen, H
AU  - Chen H
FAU - Heinsohn, H
AU  - Heinsohn H
FAU - Vandlen, R
AU  - Vandlen R
FAU - Ferrara, N
AU  - Ferrara N
LA  - eng
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Endothelial Growth Factors)
RN  - 0 (Lymphokines)
RN  - 0 (Macromolecular Substances)
RN  - 0 (Peptide Fragments)
RN  - 0 (Receptors, IgG)
RN  - 0 (Recombinant Proteins)
RN  - 0 (VEGFA protein, human)
RN  - 0 (Vascular Endothelial Growth Factor A)
RN  - 0 (Vascular Endothelial Growth Factors)
RN  - EC 3.4.- (Endopeptidases)
RN  - EC 3.4.21.7 (Fibrinolysin)
SB  - IM
MH  - Adrenal Cortex
MH  - Alternative Splicing
MH  - Animals
MH  - Binding, Competitive
MH  - CHO Cells
MH  - Capillaries
MH  - Cattle
MH  - Cell Division/drug effects
MH  - Cells, Cultured
MH  - Chromatography, Affinity
MH  - Chromatography, High Pressure Liquid
MH  - Cloning, Molecular
MH  - Cricetinae
MH  - Endopeptidases/metabolism
MH  - Endothelial Growth Factors/chemistry/*pharmacology/physiology
MH  - Endothelium, Vascular/*cytology/drug effects
MH  - Escherichia coli
MH  - Fibrinolysin/metabolism
MH  - Humans
MH  - Kinetics
MH  - Lymphokines/chemistry/*pharmacology/physiology
MH  - Macromolecular Substances
MH  - Peptide Fragments/chemistry/isolation & purification/*pharmacology
MH  - Polymorphism, Genetic
MH  - Protein Folding
MH  - Receptors, IgG/metabolism
MH  - Recombinant Proteins/chemistry/metabolism/pharmacology
MH  - Transfection
MH  - Vascular Endothelial Growth Factor A
MH  - Vascular Endothelial Growth Factors
EDAT- 1996/03/29 00:00
MHDA- 1996/03/29 00:01
CRDT- 1996/03/29 00:00
PHST- 1996/03/29 00:00 [pubmed]
PHST- 1996/03/29 00:01 [medline]
PHST- 1996/03/29 00:00 [entrez]
AID - S0021-9258(17)45193-3 [pii]
AID - 10.1074/jbc.271.13.7788 [doi]
PST - ppublish
SO  - J Biol Chem. 1996 Mar 29;271(13):7788-95. doi: 10.1074/jbc.271.13.7788.