Distinctive binding modes and inhibitory mechanisms of two peptidic inhibitors of urokinase-type plasminogen activator with isomeric P1 residues.
Two isomeric piperidine derivatives (meta and para isomers) were used as arginine mimics in the P1 position of a cyclic peptidic inhibitor (CPAYSRYLDC) of urokinase-type plasminogen activator. The two resulting cyclic peptides showed vastly different affinities (∼70 fold) to the target enzyme. X-ray crystal structure analysis showed that the two P1 residues were inserted into the S1 specificity pocket in indistinguishable manners. However, the rest of the peptides bound in entirely different ways on the surface of the enzyme, and the two peptides have different conformations, despite the highly similar sequence. These results demonstrate how the subtle difference in P1 residue can dictate the exosite interactions and the potencies of peptidic inhibitors, and highlight the importance of the P1 residue for protease inhibition. This study provides important information for the development of peptidic agents for pharmacological intervention.
Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China(1); Danish-Chinese Centre for Proteases and Cancer, China.
Longguang
L
Jiang
Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China(1); Danish-Chinese Centre for Proteases and Cancer, China.
Baoyu
B
Zhao
Department of Chemistry, Aarhus University, Aarhus, Denmark; Danish-Chinese Centre for Proteases and Cancer, China.
Peng
P
Xu
Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark(2); Danish-Chinese Centre for Proteases and Cancer, China.
Hans Peter
HP
Sørensen
Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark(2); Danish-Chinese Centre for Proteases and Cancer, China.
Jan K
JK
Jensen
Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark(2); Danish-Chinese Centre for Proteases and Cancer, China.
Anni
A
Christensen
Department of Chemistry, Aarhus University, Aarhus, Denmark; Danish-Chinese Centre for Proteases and Cancer, China.
Masood
M
Hosseini
Department of Chemistry, Aarhus University, Aarhus, Denmark; Danish-Chinese Centre for Proteases and Cancer, China.
Niels Christian
NC
Nielsen
Department of Chemistry, Faculty of Science, University of Copenhagen, Copenhagen, Denmark(3); Danish-Chinese Centre for Proteases and Cancer, China.
Knud J
KJ
Jensen
Department of Molecular Biology and Genetics, Aarhus University, Aarhus, Denmark(2); Danish-Chinese Centre for Proteases and Cancer, China.
Peter A
PA
Andreasen
Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, Fuzhou 350002, China(1); Danish-Chinese Centre for Proteases and Cancer, China. Electronic address: mhuang@fjirsm.ac.cn.
Mingdong
M
Huang
D000595:Amino Acid Sequence; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006801:Humans; D008958:Models, Molecular; D010446:Peptide Fragments / Q000737:chemistry / Q000378:metabolism / Q000494:pharmacology; D010456:Peptides, Cyclic / Q000737:chemistry / Q000494:pharmacology; D011480:Protease Inhibitors / Q000737:chemistry / Q000378:metabolism* / Q000494:pharmacology; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D014568:Urokinase-Type Plasminogen Activator / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism*
D010446:Peptide Fragments; D010456:Peptides, Cyclic; D011480:Protease Inhibitors; C530075:mupain-1; D014568:Urokinase-Type Plasminogen Activator
Cyclic peptide; Inhibitor; Protease; Urokinase; X-ray crystal structure