Functional evaluation of angiotensin convertin enzyme (ACE) by site-directed changes

Abstract

Hypertension is a worldwide health problem that is emerging in the population as a major public health challenges and considered a major risk factor for cardiovascular and renal disease. Several studies have focused on the renin-angiotensin system, which is described as an endocrine system, where renin, an aspartyl endopeptidase, cleaves angiotensinogen in the Leu10-Val11 bond and releases the decapeptide angiotensin I (AI), which is converted to angiotensin II (AII), by the action of angiotensin-converting enzyme (ACE). ACE is an enzyme target of several therapeutic studies, and thus, the search for better understanding of its structure is the goal of many researchers. Thus, this study aims to investigate the general role of amino acid residues in the ACE inhibitor lisinopril interaction with the site located in the N-terminal protein shown during the studies of the structure of the N-domain isoforms of ACE, as well as the role of these amino acids on the stability and functionality of the enzyme. Thus, this project targets the residues are Asp140, Gln259, His331, Ala332, Ser333, Gln355, Thr358 and Phe435 ACE, strong candidates for interaction with the ACE inhibitor lisinopril. In addition, the Arg500 residue will also be reviewed, since it appears to be essential for the binding of chloride ion. It is expected that mutations in these positions may compromise the interaction of the ligand (substrate / inhibitor) with the ACE or even change the way this interaction, making it unproductive. So they want to mutate amino acid residues to alanine and analyze the effect of the mutation made in relation to the processes of ligand-enzyme interaction and functional activity of the enzyme. The residue Ala332 is mutated to tryptophan space to verify the importance of this residue in the enzyme. In addition, we intend to analyze the importance of these amino acid residues to the newly described role of ACE as a molecule able to transduce signal and their importance for the N-domain isoform of ACE 90kDa possible genetic marker of hypertension. (AU)