PMID- 10713063
OWN - NLM
STAT- MEDLINE
DCOM- 20000412
LR  - 20231213
IS  - 0021-9258 (Print)
IS  - 0021-9258 (Linking)
VI  - 275
IP  - 11
DP  - 2000 Mar 17
TI  - Recombinant toxins that bind to the urokinase receptor are cytotoxic without 
      requiring binding to the alpha(2)-macroglobulin receptor.
PG  - 7566-73
AB  - The alpha(2-)macroglobulin receptor (alpha(2)MR) has been reported to mediate the 
      internalization of the urokinase plasminogen activator receptor (uPAR) via ligand 
      binding to both receptors. To target malignant uPAR-expressing cells and to 
      determine whether uPAR can internalize without ligand binding to alpha(2)MR, we 
      engineered two recombinant toxins, ATF-PE38 and ATF-PE38KDEL. Each consists of 
      the amino-terminal fragment (ATF) of human urokinase and a truncated form of 
      Pseudomonas exotoxin (PE) devoid of domain Ia, which binds alpha(2)MR. ATF-PE38 
      and ATF-PE38KDEL were cytotoxic toward malignant uPAR-bearing cells, with IC(50) 
      values as low as 0.02 ng/ml (0.3 pM). Cytotoxicity could be blocked using either 
      recombinant urokinase or free ATF, indicating that the cytotoxicity of the 
      recombinant toxins was specific. Radiolabeled ATF-PE38 had high affinity for uPAR 
      (K(d) = 0.4-8 nM) on a variety of different malignant cell types and internalized 
      at a rate similar to that of ATF. The cytotoxicity was not diminished by 
      receptor-associated protein, which binds and shields the alpha(2)MR from other 
      proteins, or by incubation with phorbol myristate acetate, which is known to 
      decrease the number of alpha(2)MRs in U937 cells or by antibodies to alpha(2)MR. 
      Therefore, these recombinant toxins appear to internalize via uPAR without 
      association with the alpha(2)MR.
FAU - Rajagopal, V
AU  - Rajagopal V
AD  - Laboratory of Molecular Biology, Division of Basic Sciences, NCI, National 
      Institutes of Health, Bethesda, Maryland 20892, USA.
FAU - Kreitman, R J
AU  - Kreitman RJ
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PL  - United States
TA  - J Biol Chem
JT  - The Journal of biological chemistry
JID - 2985121R
RN  - 0 (Antineoplastic Agents)
RN  - 0 (Bacterial Toxins)
RN  - 0 (Exotoxins)
RN  - 0 (Immunotoxins)
RN  - 0 (Low Density Lipoprotein Receptor-Related Protein-1)
RN  - 0 (PLAUR protein, human)
RN  - 0 (Peptide Fragments)
RN  - 0 (Receptors, Cell Surface)
RN  - 0 (Receptors, Immunologic)
RN  - 0 (Receptors, Urokinase Plasminogen Activator)
RN  - 0 (Recombinant Proteins)
RN  - 0 (Virulence Factors)
RN  - EC 2.4.2.- (ADP Ribose Transferases)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
RN  - NI40JAQ945 (Tetradecanoylphorbol Acetate)
SB  - IM
MH  - *ADP Ribose Transferases
MH  - Antineoplastic Agents/metabolism/pharmacology
MH  - Bacterial Toxins/genetics/metabolism/pharmacology
MH  - Binding Sites
MH  - Binding, Competitive
MH  - Biological Transport
MH  - Drug Stability
MH  - Exotoxins/genetics/*metabolism/pharmacology
MH  - Humans
MH  - Immunotoxins/genetics/*metabolism/pharmacology
MH  - Low Density Lipoprotein Receptor-Related Protein-1
MH  - Lymphocyte Activation
MH  - Peptide Fragments/genetics/metabolism/pharmacology
MH  - Protein Binding
MH  - Pseudomonas aeruginosa
MH  - Receptors, Cell Surface/*metabolism
MH  - Receptors, Immunologic/*metabolism
MH  - Receptors, Urokinase Plasminogen Activator
MH  - Recombinant Proteins/metabolism
MH  - Structure-Activity Relationship
MH  - Tetradecanoylphorbol Acetate/pharmacology
MH  - Toxicity Tests
MH  - Tumor Cells, Cultured/drug effects
MH  - U937 Cells
MH  - Urokinase-Type Plasminogen Activator/genetics/*metabolism/pharmacology
MH  - *Virulence Factors
MH  - Pseudomonas aeruginosa Exotoxin A
EDAT- 2000/03/14 09:00
MHDA- 2000/04/15 09:00
CRDT- 2000/03/14 09:00
PHST- 2000/03/14 09:00 [pubmed]
PHST- 2000/04/15 09:00 [medline]
PHST- 2000/03/14 09:00 [entrez]
AID - S0021-9258(18)30250-3 [pii]
AID - 10.1074/jbc.275.11.7566 [doi]
PST - ppublish
SO  - J Biol Chem. 2000 Mar 17;275(11):7566-73. doi: 10.1074/jbc.275.11.7566.