PMID- 1510944
OWN - NLM
STAT- MEDLINE
DCOM- 19920929
LR  - 20190613
IS  - 0006-2960 (Print)
IS  - 0006-2960 (Linking)
VI  - 31
IP  - 33
DP  - 1992 Aug 25
TI  - Heparin binding to the urokinase kringle domain.
PG  - 7572-9
AB  - The binding of urokinase to immobilized heparin and dextran sulfate was studied 
      using activity assays of the bound urokinase. The markedly higher binding 
      observed with high M(r) urokinase compared to low M(r) urokinase indicated a role 
      for the amino-terminal fragment (ATF). This was confirmed by the use of inactive 
      truncated urokinase and monoclonal antibodies specific for the ATF in competition 
      assays of urokinase binding. Antibody competition assays suggested a site in the 
      kringle domain, and a synthetic decapeptide Arg-52-Trp-62 from the kringle 
      sequence (kringle numbering convention) was competitive in assays of urokinase 
      binding to dextran sulfate and heparin. Heparin binding to the urokinase kringle 
      was unambiguously demonstrated via 1H NMR spectroscopy at 500 MHz. Effective 
      equilibrium association constants (K(a)*) were determined for the interaction of 
      isolated kringle fragment and low M(r) heparin at pH 7.2. The binding was strong 
      in salt-free 2H2O (K(a)* approximately 57 mM-1) and remained significant in 0.15 
      M NaCl (K(a)* approximately 12 mM-1), supporting a potential physiological role 
      for the interaction. This is the first demonstration of a function for the 
      kringle domain of urokinase, and it suggests that while the classical kringle 
      structure has specificity for lysine binding, there may also exist a class of 
      kringles with affinity for polyanion binding.
FAU - Stephens, R W
AU  - Stephens RW
AD  - Department of Virology, University of Helsinki, Finland.
FAU - Bokman, A M
AU  - Bokman AM
FAU - Myohanen, H T
AU  - Myohanen HT
FAU - Reisberg, T
AU  - Reisberg T
FAU - Tapiovaara, H
AU  - Tapiovaara H
FAU - Pedersen, N
AU  - Pedersen N
FAU - Grondahl-Hansen, J
AU  - Grondahl-Hansen J
FAU - Llinas, M
AU  - Llinas M
FAU - Vaheri, A
AU  - Vaheri A
LA  - eng
GR  - HL-29409/HL/NHLBI 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.
PL  - United States
TA  - Biochemistry
JT  - Biochemistry
JID - 0370623
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Peptide Fragments)
RN  - 9005-49-6 (Heparin)
RN  - 9042-14-2 (Dextran Sulfate)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Antibodies, Monoclonal
MH  - Binding Sites
MH  - Cell Line
MH  - Chromatography, Affinity
MH  - Dextran Sulfate/metabolism
MH  - Heparin/*metabolism
MH  - Humans
MH  - Kinetics
MH  - Magnetic Resonance Spectroscopy
MH  - Mice
MH  - Molecular Sequence Data
MH  - Peptide Fragments/chemistry/metabolism
MH  - Protein Conformation
MH  - Transfection
MH  - Urokinase-Type Plasminogen Activator/chemistry/genetics/*metabolism
EDAT- 1992/08/25 00:00
MHDA- 1992/08/25 00:01
CRDT- 1992/08/25 00:00
PHST- 1992/08/25 00:00 [pubmed]
PHST- 1992/08/25 00:01 [medline]
PHST- 1992/08/25 00:00 [entrez]
AID - 10.1021/bi00148a019 [doi]
PST - ppublish
SO  - Biochemistry. 1992 Aug 25;31(33):7572-9. doi: 10.1021/bi00148a019.