PDBsum entry 1vj9

Hydrolase

PDB id: 1vj9

Contents

Protein chain

247 a.a. *

Ligands

SO4 ×2

5IN

Waters ×92

* Residue conservation analysis

PDB id:

1vj9

Name:

Hydrolase

Title:

Urokinase plasminogen activator b-chain-jt464 complex

Structure:

Plasminogen activator, urokinase. Chain: u. Fragment: b chain. Engineered: yes. Mutation: yes

Source:

Homo sapiens. Human. Organism_taxid: 9606. Expressed in: escherichia coli. Expression_system_taxid: 562.

Resolution:

2.40Å R-factor: not given R-free: 0.247

Authors:

A.Schweinitz,T.Steinmetzer,I.J.Banke,M.J.E.Arlt,A.Stuerzebecher, O.Schuster,A.Geissler,H.Giersiefen,E.Zeslawska,U.Jacob,A.Kruger, J.Stuerzebecher

Key ref:

A.Schweinitz et al. (2004). Design of novel and selective inhibitors of urokinase-type plasminogen activator with improved pharmacokinetic properties for use as antimetastatic agents. J Biol Chem, 279, 33613-33622. PubMed id: 15150279 DOI: 10.1074/jbc.M314151200

Date:

03-Feb-04 Release date: 22-Jun-04

Protein chain

P00749  (UROK_HUMAN) -  Urokinase-type plasminogen activator from Homo sapiens

Seq: 431 a.a.

Struc: 247 a.a.*

* PDB and UniProt seqs differ at 1 residue position (black cross)

Enzyme reactions

• Enzyme class: E.C.3.4.21.73 - u-plasminogen activator.
• Reaction: Specific cleavage of Arg-|-Val bond in plasminogen to form plasmin.

DOI no: 10.1074/jbc.M314151200

J Biol Chem 279:33613-33622 (2004)

PubMed id: 15150279

Design of novel and selective inhibitors of urokinase-type plasminogen activator with improved pharmacokinetic properties for use as antimetastatic agents.

A.Schweinitz, T.Steinmetzer, I.J.Banke, M.J.Arlt, A.Stürzebecher, O.Schuster, A.Geissler, H.Giersiefen, E.Zeslawska, U.Jacob, A.Krüger, J.Stürzebecher.

ABSTRACT

The serine protease urokinase-type plasminogen activator (uPA) interacts with a specific receptor (uPAR) on the surface of various cell types, including tumor cells, and plays a crucial role in pericellular proteolysis. High levels of uPA and uPAR often correlate with poor prognosis of cancer patients. Therefore, the specific inhibition of uPA with small molecule active-site inhibitors is one strategy to decrease the invasive and metastatic activity of tumor cells. We have developed a series of highly potent and selective uPA inhibitors with a C-terminal 4-amidinobenzylamide residue. Optimization was directed toward reducing the fast elimination from circulation that was observed with initial analogues. The x-ray structures of three inhibitor/uPA complexes have been solved and were used to improve the inhibition efficacy. One of the most potent and selective derivatives, benzylsulfonyl-D-Ser-Ser-4-amidinobenzylamide (inhibitor 26), inhibits uPA with a Ki of 20 nm. This inhibitor was used in a fibrosarcoma model in nude mice using lacZ-tagged human HT1080 cells, to prevent experimental lung metastasis formation. Compared with control (100%), an inhibitor dose of 2 x 1.5 mg/kg/day reduced the number of experimental metastases to 4.6 +/- 1%. Under these conditions inhibitor 26 also significantly prolonged survival. All mice from the control group died within 43 days after tumor cell inoculation, whereas 50% of mice from the inhibitor-treated group survived more than 117 days. This study demonstrates that the specific inhibition of uPA by these inhibitors may be a useful strategy for the treatment of cancer to prevent metastasis.

Selected figure(s)

Figure 2.
FIG. 2. Stereo view of the active site region of c-uPA in complex with inhibitors 1 (A, Protein Data Bank code 1SC8 [PDB] ) and 26 (B, Protein Data Bank code 1VJA [PDB] ). uPA and inhibitor residues are drawn with yellow and white carbon atoms, respectively. Characteristic hydrogen bonds are shown as thin white lines, selected amino acid residues of c-uPA are labeled.

Figure 4.
FIG. 4. Stereo view of the model of the inhibitor 34·human-uPA complex. The inhibitor is drawn as sticks with atom dependent colors. For simplification, only the hydrogens, which are attached to the terminal side chain nitrogens of the P2 arginine, are shown. The protein is visualized by a Connolly surface, blue and red surface areas showing hydrogen acceptors and donators, respectively. Gray areas have no hydrogen bonding properties. The guanidino group of the Arg side chain in the P2 position of the inhibitor probably forms a salt bridge (yellow lines with distances given in Å) to the carboxyl group of Asp60A, found specifically only in human uPA. This model was generated from the x-ray structure of the inhibitor 27· c-uPA complex (see Fig. 3) by replacement of the oxygen and phenyl ring in the Ser(Bzl) side chain by a CH[2]- and guanidino group, respectively, followed by energy minimization of the enzyme-inhibitor complex using the software package Sybyl version 6.9.1. (Tripos).

The above figures are reprinted by permission from the ASBMB: J Biol Chem (2004, 279, 33613-33622) copyright 2004.

Figures were selected by an automated process.