PDBsum entry 4zhm

Hydrolase inhibitor/hydrolase

PDB id: 4zhm

Contents

Protein chain

247 a.a.

Ligands

CYS-PRO-ALA-TYR-SER-ALA-TYR-ILE-GLY-CYS

MRZ

Waters ×57

PDB id:

4zhm

Name:

Hydrolase inhibitor/hydrolase

Title:

The crystal structure of mupain-1--16-ig in complex with murinised human upa at ph7.4

Structure:

Mupain-1-16-ig. Chain: p. Engineered: yes. Urokinase-type plasminogen activator. Chain: u. Fragment: unp residues 179-425. Synonym: upa. Engineered: yes. Mutation: yes

Source:

Synthetic: yes. Synthetic construct. Organism_taxid: 32630. Homo sapiens. Human. Organism_taxid: 9606. Gene: plau. Expressed in: komagataella pastoris. Expression_system_taxid: 4922

Resolution:

1.90Å R-factor: 0.226 R-free: 0.264

Authors:

L.Jiang,P.A.Andreasen,M.Huang

Key ref:

H.P.Sørensen et al. (2015). Selection of High-Affinity Peptidic Serine Protease Inhibitors with Increased Binding Entropy from a Back-Flip Library of Peptide-Protease Fusions. J Mol Biol, 427, 3110-3122. PubMed id: 26281711 DOI: 10.1016/j.jmb.2015.08.005

Date:

25-Apr-15 Release date: 16-Sep-15

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 3 residue positions (black crosses)

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: 10.1016/j.jmb.2015.08.005

J Mol Biol 427:3110-3122 (2015)

PubMed id: 26281711

Selection of High-Affinity Peptidic Serine Protease Inhibitors with Increased Binding Entropy from a Back-Flip Library of Peptide-Protease Fusions.

H.P.Sørensen, P.Xu, L.Jiang, T.Kromann-Hansen, K.J.Jensen, M.Huang, P.A.Andreasen.

ABSTRACT

We have developed a new concept for designing peptidic protein modulators, by recombinantly fusing the peptidic modulator, with randomized residues, directly to the target protein via a linker and screening for internal modulation of the activity of the protein. We tested the feasibility of the concept by fusing a 10-residue-long, disulfide-bond-constrained inhibitory peptide, randomized in selected positions, to the catalytic domain of the serine protease murine urokinase-type plasminogen activator. High-affinity inhibitory peptide variants were identified as those that conferred to the fusion protease the lowest activity for substrate hydrolysis. The usefulness of the strategy was demonstrated by the selection of peptidic inhibitors of murine urokinase-type plasminogen activator with a low nanomolar affinity. The high affinity could not have been predicted by rational considerations, as the high affinity was associated with a loss of polar interactions and an increased binding entropy.