Simulated interactions between angiotensin-converting enzyme and substrate gonadotropin-releasing hormone: novel insights into domain selectivity.
Human angiotensin-I converting enzyme (ACE) is a central component of the renin-angiotensin system and a major target for cardiovascular therapies. The somatic form of the enzyme (sACE) comprises two homologous metallopeptidase domains (N and C), each bearing a zinc active site with similar but distinct substrate and inhibitor specificities. On the basis of the recently determined crystal structures of both ACE domains, we have studied their complexes with gonadotropin-releasing hormone (GnRH), which is cleaved releasing both the protected NH2- and COOH-terminal tripeptides. This is the first molecular modeling study of an ACE-peptide substrate complex that examines the structural basis of ACE's endopeptidase activity and offers novel insights into subsites that are distant from the obligatory binding site and were not identified in the crystal structures. Our data indicate that a bridging interaction between Arg500 of the N-domain and Arg8 of GnRH that involves a buried chloride ion may account for its role in the specificity of the N-domain for endoproteolytic cleavage of the substrate at the NH2-terminus in vitro. In support of this, the protected NH2-terminal dipeptide of GnRH exhibits stronger interactions than the protected COOH-terminal dipeptide with the N-domain of ACE. Further comparison of the models of ACE-substrate complexes promotes our understanding of how the two domains differ in their function and specificity and provides an extension of the pharmacophore model used for structure-based drug design up to the S7 subsite of the enzyme.
Papakyriakou
Athanasios
A
Department of Pharmacy, University of Patras, Panepistimioupoli-Rion, GR-26504, Greece.
Spyroulias
Georgios A
GA
Sturrock
Edward D
ED
Manessi-Zoupa
Evy
E
Cordopatis
Paul
P
D020816:Amino Acid Motifs; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000378:metabolism; D001665:Binding Sites; D020134:Catalytic Domain; D003198:Computer Simulation; D018360:Crystallography, X-Ray; D020689:Exopeptidases / Q000378:metabolism; D007987:Gonadotropin-Releasing Hormone / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D045726:Metalloproteases / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D012965:Sodium Chloride / Q000737:chemistry / Q000378:metabolism; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D015032:Zinc / Q000737:chemistry
D000806:Angiotensin-Converting Enzyme Inhibitors; D007987:Gonadotropin-Releasing Hormone; D012965:Sodium Chloride; D020689:Exopeptidases; D045726:Metalloproteases; D007703:Peptidyl-Dipeptidase A; D015032:Zinc