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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

New approach for natural products screening by real-time monitoring of hemoglobin hydrolysis using quartz crystal microbalance

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Author(s):
Cornelio, Vivian E. [1] ; Pedroso, Mariele M. [1] ; Afonso, Andre S. [1] ; Fernandes, Joao B. [1] ; da Silva, M. Fatima G. F. [1] ; Faria, Ronaldo C. [1] ; Vieira, Paulo C. [1]
Total Authors: 7
Affiliation:
[1] Fed Univ Sao Carlos UFSCar, Dept Chem, Sao Carlos, SP - Brazil
Total Affiliations: 1
Document type: Journal article
Source: Analytica Chimica Acta; v. 862, p. 86-93, MAR 3 2015.
Web of Science Citations: 5
Abstract

The hemoglobin (Hb) released from erythrocytes is a primary nutritive component for many blood-feeding parasites. The aspartic protease cathepsin D is a hemoglobinase that is involved in the Hb degradation process and is considered an interesting target for chemotherapy intervention. However, traditional enzymatic assays for studying Hb degradation utilize spectrophotometric techniques, which do not allow real-time monitoring and can present serious interference problems. Herein, we describe a biosensor using simple approach for the real-time monitoring of Hb hydrolysis as well as an efficient screening method for natural products as enzymatic inhibitors using a quartz crystal microbalance (QCM) technique. Hemoglobin was anchored on the quartz crystal surface using mixed self-assembled monolayers. The addition of the enzyme caused a mass change (frequency shift) due to Hb hydrolysis, which was monitored in real time. From the frequency change patterns of the Hb-functionalized QCM, we evaluated the enzymatic reaction by determining the kinetic parameters of product formation (k(cat)). The QCM enzymatic assay using immobilized human Hb was shown to be an excellent approach for screening possible inhibitors in complex mixtures, opening up a new avenue for the discovery of novel inhibitors. (C) 2015 Elsevier B.V. All rights reserved. (AU)

FAPESP's process: 09/17538-8 - Study of new methodologies for cathepsin D assays in the search for inhibitors
Grantee:Vivian Estevam Cornélio
Support Opportunities: Scholarships in Brazil - Doctorate (Direct)