Exploring Enzymatic Conformational Dynamics at Surfaces through μ-FTIR Spectromicroscopy

Author(s)

Mendes, Giovana Rossi ; Modenez, Iago de Assis ; Cagnani, Giovana Rosso ; Colombo, Rafael N. P. ; Crespilho, Frank Nelson

• Document type: Journal article
• Source: Analytical Chemistry; v. 95, n. 30, p. 9-pg., 2023-07-17.

Abstract

Immobilization of proteins onto solidsupports has criticalindustrial,technological, and medical applications, and is a daily task in chemicalresearch. Significant conformational rearrangements often occur dueto enzyme-surface interactions, and it is of broad interestto develop methods to probe and better understand these molecular-levelchanges that contribute to the enzyme's catalytic activityand stability. While circular dichroism is a common method for solution-phaseconformational study, the application to surface-supported proteinsis not trivial and spatial mapping is not viable. On the other hand,a nonlinear laser spectroscopy technique used to analyze surfacesand interfaces is not often found in most laboratories, thereforerequiring an alternative and reliable method. Here, we employed high-dimensionaldata spectromicroscopy analysis in the infrared region (& mu;-FTIR)to investigate the enzyme's conformational change when adsorbedonto solid matrices, across a ca. 20 mm(2) area. Alcoholdehydrogenase (ADH) enzyme was adopted as a model enzyme to interactwith CaF2, Au, and Au-thiol model substrates, strategicallychosen for mapping the enzyme dynamics on solid surfaces with differentpolarity/hydrophobicity properties and extendable to other materials.Two-dimensional chemical maps indicate that the enzyme adsorbs withdifferent patterns in which secondary structures dynamically adjustto optimize interprotein and enzyme-surface interactions. Theresults suggest an experimental approach to identify and map enzymeconformational dynamics onto different solid surfaces across spaceand provide insights into immobilized protein structure investigationsfor areas such as biosensing and bioenergy. (AU)

• FAPESP's process: 20/10724-0 - Biomolecules dynamic in solid surfaces using spectromicroscopy
• Grantee: Giovana Rossi Mendes
• Support Opportunities: Scholarships in Brazil - Scientific Initiation
• FAPESP's process: 21/05665-7 - Development of Van der Waals surfaces for application in biodevices
• Grantee: Rafael Neri Prystaj Colombo
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 19/15333-1 - Bio-photo-electrochemical hybrid cells for solar energy conversion
• Grantee: Frank Nelson Crespilho
• Support Opportunities: Regular Research Grants
• FAPESP's process: 19/12053-8 - High performance electrodes applied in organic batteries and in biofuel cell
• Grantee: Frank Nelson Crespilho
• Support Opportunities: Regular Research Grants
• FAPESP's process: 18/22214-6 - Towards a convergence of technologies: from sensing and biosensing to information visualization and machine learning for data analysis in clinical diagnosis
• Grantee: Osvaldo Novais de Oliveira Junior
• Support Opportunities: Research Projects - Thematic Grants
• FAPESP's process: 22/09164-5 - Understanding the charge transfer mechanisms in biological and biomimetic systems using in-situ and operando techniques: from fundamental science to energy application
• Grantee: Frank Nelson Crespilho
• Support Opportunities: Regular Research Grants