Angiotensin I-converting enzyme transition state stabilization by HIS1089 : evidence for a catalytic mechanism distinct from other gluzincin metalloproteinases

Angiotensin (Ang) I-converting enzyme (ACE) is a member of the gluzincin family of zinc metalloproteinases that contains two homologous catalytic domains. Both the N- and C-terminal domains are peptidyl-dipeptidases that catalyze Ang II formation and bradykinin degradation. Multiple sequence alignment was used to predict His¹⁰⁸⁹ as the catalytic residue in human ACE C-domain that, by analogy with the prototypical gluzincin, thermolysin, stabilizes the scissile carbonyl bond through a hydrogen bond during transition state binding. Site-directed mutagenesis was used to change His¹⁰⁸⁹ to Ala or Leu. At pH 7.5, with Ang I as substrate, k_cat/K_m values for these Ala and Leu mutants were 430 and 4,000-fold lower, respectively, compared with wild-type enzyme and were mainly due to a decrease in catalytic rate (k_cat) with minor effects on ground state substrate binding (K_m). A 120,000-fold decrease in the binding of lisinopril, a proposed transition state mimic, was also observed with the His¹⁰⁸⁹ --> Ala mutation. ACE C-domain-dependent cleavage of AcAFAA showed a pH optimum of 8.2. H1089A has a pH optimum of 5.5 with no pH dependence of its catalytic activity in the range 6.5-10.5, indicating that the His¹⁰⁸⁹ side chain allows ACE to function as an alkaline peptidyl-dipeptidase. Since transition state mutants of other gluzincins show pH optima shifts toward the alkaline, this effect of His¹⁰⁸⁹ on the ACE pH optimum and its ability to influence transition state binding of the sulfhydryl inhibitor captopril indicate that the catalytic mechanism of ACE is distinct from that of other gluzincins.

The domain layer for mixed-initiative interaction in generative design

The design space exploration formalism has developed data structures and algorithms of sufficient complexity and scope to support conceptual layout, massing, and enclosure configurations. However, design remains a human enterprise. To support the user in designing with the formalism, we have developed an interaction model that addresses the interleaving of user actions with the formal operations of design space exploration. The central feature of our interaction model is the modeling of control based on mixed-initiative. Initiative is sometimes taken by the designer and sometimes by the formalism in working on a shared design task. The model comprises three layers, domain, task, and dialogue. In this paper we describe the formulation of the domain layer of our mixed-initiative interaction model for design space exploration. We present the view of the domain as understood in the formalism in terms of the three abstract concepts of state, move, and structure. In order to support mixed initiative, it is necessary to develop a shared view of the domain. The domain layer addresses this problem by mapping the designer's view onto the symbol substrate. First, we present the designer's view of the domain in terms of problems, solutions, choices, and history. Second, we show how this view is interleaved with the symbol-substrate through four domain layer constructs, problem state, solution state, choice, and exploration history. The domain layer presents a suitable foundation for integrating the role of the designer with a description formalism. It enables the designer to maintain exploration freedom in terms of formulating and reformulating problems, generating solutions, making choices, and navigating the history of exploration.

Half metallicity in a zigzag double-walled nanotube nanodot : an ab initio prediction

Ab initio density functional calculations were performed to study finite-length zigzag (7, 0) @ (16, 0) double-walled carbon nanotubes (DWCNTs) with H-termination at the open ends. We find that such a DWCNT nanodot displays a very large magnetic moment at the zigzag edges and the ground state displays symmetric anti-ferromagnetic coupling. When an external electric field is applied along the direction of tube axis, a gap is opened for one spin channel, whereas another spin channel remains metallic, i.e. half metallicity occurs. Our results suggest an important new avenue for the development of CNT-based spintronic materials with enhanced properties. © 2008 Elsevier B.V. All rights reserved.