title: Molecular basis of exopeptidase activity in the C-terminal domain of human angiotensin I-converting enzyme: insights into the origins of its exopeptidase activity.
abstract: Proteolytic processing is a primary means of biological control. Exopeptidases use terminal anchoring interactions to restrict cleavage at peptide substrate N or C termini. In contrast, internal peptide bond targeting by endopeptidases is through context-driven recognition. Angiotensin I-converting enzyme (ACE), a zinc metalloproteinase, has tandem duplicate catalytic domains, N- and C-terminal, each of which is a dual specificity enzyme with exo- and endocarboxypeptidase activities. The mechanisms by which ACE evolved from its endopeptidase ancestors as a dual specificity enzyme have not been defined. Based on kinetic studies of wild-type and mutant forms of the C-terminal catalytic domain of human ACE and of the ACE substrates angiotensin I, substance P, and bradykinin, as well as considerations of the ACE x-ray structure, we provide evidence that the acquisition of its exopeptidase activity is due to novel evolutionary specializations. These involve not only interactions between the S(2)' subsite cognate for the C-terminal substrate P(2)' side chain, acting in concert with carboxylate-docking interactions with Lys(1087) and Tyr(1096), but also electrostatic selection against a cationic C-terminal substrate carboxylate. With a blocked C terminus, substrate side chain interactions are dominant in cleavage site selection. In the evolution of obligate exopeptidases from endopeptidase ancestors, mutations that destroy context-driven peptide bond targeting are likely to have followed the acquisition of terminal docking interactions. Evolutionary intermediates between endopeptidases and obligate exopeptidases could therefore have been dual specificity proteinases like ACE.
authors.affiliation: Victor Chang Cardiac Research Institute, Darlinghurst, New South Wales 2010, Australia.
authors.forename: Nawazish
authors.initials: N
authors.lastname: Naqvi
authors.forename: Ke
authors.initials: K
authors.lastname: Liu
authors.forename: Robert M
authors.initials: RM
authors.lastname: Graham
authors.forename: Ahsan
authors.initials: A
authors.lastname: Husain
mesh_terms: D000803:Angiotensin I / Q000378:metabolism; D001920:Bradykinin / Q000378:metabolism; D003198:Computer Simulation; D019143:Evolution, Molecular; D020689:Exopeptidases / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D009154:Mutation; D007703:Peptidyl-Dipeptidase A / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D055672:Static Electricity; D013373:Substance P / Q000378:metabolism; D013379:Substrate Specificity
chemical_list: D013373:Substance P; D000803:Angiotensin I; D020689:Exopeptidases; D007703:Peptidyl-Dipeptidase A; D001920:Bradykinin