PMID- 10446189
OWN - NLM
STAT- MEDLINE
DCOM- 19990909
LR  - 20190508
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 274
IP  - 34
DP  - 1999 Aug 20
TI  - Human heparanase. Purification, characterization, cloning, and expression.
PG  - 24153-60
AB  - Heparan sulfate and heparan sulfate proteoglycans are present in the
      extracellular matrix as well as on the external cell surface. They bind various
      molecules such as growth factors and cytokines and modulate the biological
      functions of binding proteins. Heparan sulfate proteoglycans are also important
      structural components of the basement membrane. Heparanase is an
      endo-beta-D-glucuronidase capable of cleaving heparan sulfate and has been
      implicated in inflammation and tumor angiogenesis and metastasis. In this study, 
      we report the purification of a human heparanase from an SV40-transformed
      embryonic fibroblast cell line WI38/VA13 by four sequential column
      chromatographies. The activity was measured by high speed gel permeation
      chromatography of the degradation products of fluorescein isothiocyanate-labeled 
      heparan sulfate. The enzyme was purified to homogeneity, yielding a peptide with 
      an apparent molecular mass of 50 kDa when analyzed by SDS-polyacrylamide gel
      electrophoresis. Using the amino acid sequences of the N-terminal and internal
      heparanase peptides, a cDNA coding for human heparanase was cloned. NIH3T3 and
      COS-7 cells stably transfected with pBK-CMV expression vectors containing the
      heparanase cDNA showed high heparanase activities. The homology search revealed
      that no homologous protein had been reported.
FAU - Toyoshima, M
AU  - Toyoshima M
AD  - Discovery Research, Takarazuka Research Institute, Novartis Pharma K. K., 10-66
      Miyuki-cho, Takarazuka 665-8666, Japan. motowa.nakajima@pharma.novartis.com
FAU - Nakajima, M
AU  - Nakajima M
LA  - eng
SI  - GENBANK/AF155510
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - EC 3.2.1.- (Glycoside Hydrolases)
RN  - EC 3.2.1.- (heparanase)
RN  - EC 3.2.1.31 (Glucuronidase)
RN  - I223NX31W9 (Fluorescein-5-isothiocyanate)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - COS Cells
MH  - Cloning, Molecular
MH  - Fluorescein-5-isothiocyanate/metabolism
MH  - *Glucuronidase
MH  - Glycoside Hydrolases/chemistry/genetics/*isolation & purification
MH  - Humans
MH  - Hydrogen-Ion Concentration
MH  - Molecular Sequence Data
EDAT- 1999/08/14 00:00
MHDA- 1999/08/14 00:01
CRDT- 1999/08/14 00:00
PHST- 1999/08/14 00:00 [pubmed]
PHST- 1999/08/14 00:01 [medline]
PHST- 1999/08/14 00:00 [entrez]
AID - 10.1074/jbc.274.34.24153 [doi]
PST - ppublish
SO  - J Biol Chem. 1999 Aug 20;274(34):24153-60. doi: 10.1074/jbc.274.34.24153.