A three-dimensional construction of the active site (region 507749) of human neutral endopeptidase (EC.3.4.24.11)

A three-dimensional model of the 507–749 region of neutralendopeptidase-24.11 (NEP; E.C.3.4.24.11) was constructed integratingthe results of secondary structure predictions and sequencehomologies with the bacterial endopeptidase thermolysin. Additionaldata were extracted from the structure of two other metalloproteases,astacin and stromelysin. The resulting model accounts for themain biological properties of NEP and has been used to describethe environment close to the zinc atom defining the catalyticsite. The analysis of several thiol inhibitors, complexed inthe model active site, revealed the presence of a large hydrophobicpocket at the S₁' subsite level. This is supported by the natureof the constitutive amino acids. The computed energies of boundinhibitors correspond with the relative affinities of the stereoisomersof benzofused macrocycle derivatives of thiorphan. The modelcould be used to facilitate the design of new NEP inhibitors,as illustrated in the paper.