Rational structure-based design of a novel carboxypeptidase R inhibitor.
A novel carboxypeptidase R (CPR) inhibitor, related to potato carboxypeptidase inhibitor (PCI), was designed using rational structure-based strategies, incorporating two principle facts: CPR has a strong affinity for basic amino acids, and the two lysine and arginine residues of PCI are orientated in the same direction and held in close spatial proximity by three disulfide bonds. Initially, a disulfide-bonded fragment of PCI was synthesized showing weak competitive inhibitory activity against CPR. Subsequently, a smaller linear 9-mer peptide, designated CPI-2KR, was designed/synthesized and found to be a more efficient competitive inhibitor of CPR, without affecting the activity of the other plasma carboxypeptidase, carboxypeptidase N. In vitro studies showed that, together with tissue plasminogen activator, CPI-2KR synergistically accelerated fibrinolysis, representing a lead compound for the design of smaller organic molecules for use in thrombolytic therapy.
Lazoura
Eliada
E
Department of Molecular Biology, School of Medicine, Nagoya City University, Mizuho-ku, Nagoya 467-8601, Japan.
Campbell
William
W
Yamaguchi
Yoshiki
Y
Kato
Koichi
K
Okada
Noriko
N
Okada
Hidechika
H
D000595:Amino Acid Sequence; D001120:Arginine / Q000737:chemistry; D001667:Binding, Competitive; D025901:Carboxypeptidase B2 / Q000037:antagonists & inhibitors* / Q000737:chemistry* / Q000378:metabolism; D002942:Circular Dichroism; D019281:Dimerization; D004305:Dose-Response Relationship, Drug; D015195:Drug Design; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D005342:Fibrinolysis; D007700:Kinetics; D008239:Lysine / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D009842:Oligopeptides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D010940:Plant Proteins / Q000235:genetics; D011480:Protease Inhibitors; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
D004791:Enzyme Inhibitors; D009842:Oligopeptides; D010940:Plant Proteins; D011480:Protease Inhibitors; D001120:Arginine; D025901:Carboxypeptidase B2; D008239:Lysine