Paolo Mereghetti1, Paola Antonia Corsetto2, Andrea Cremona2, Angela Maria Rizzo2, Silvia Maria Doglia3, Diletta Ami4. 1. Center for Nanotechnology Innovation @NEST, Italian Institute of Technology, Piazza San Silvestro 12, Pisa 56127, Italy. 2. Department of Pharmacological and Biomolecular Sciences, BBFI, Università degli Studi di Milano, via D. Trentacoste 2, 20134 Milano, Italy. 3. Department of Physics, University of Milano-Bicocca, Piazza della Scienza 3, Milano 20126, Italy; Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milano 20126, Italy; Consorzio Nazionale Interuniversitario per le Scienze fisiche della Materia (CNISM) UdR Milano-Bicocca, Milano 20126, Italy. 4. Department of Physics, University of Milano-Bicocca, Piazza della Scienza 3, Milano 20126, Italy; Department of Biotechnology and Biosciences, University of Milano-Bicocca, Piazza della Scienza 2, Milano 20126, Italy; Consorzio Nazionale Interuniversitario per le Scienze fisiche della Materia (CNISM) UdR Milano-Bicocca, Milano 20126, Italy. Electronic address: diletta.ami@unimib.it.
Abstract
BACKGROUND: Detergent resistant membranes (DRMs) are a useful model system for the in vitro characterization of cell membrane domains. Indeed, DRMs provide a simple model to study the mechanisms underlying several key cell processes based on the interplay between specific cell membrane domains on one hand, and specific proteins and/or lipids on the other. Considering therefore their biological relevance, the development of methods capable to provide information on the composition and structure of membrane domains and to detect their modifications is highly desirable. In particular, Fourier transform infrared (FTIR) spectroscopy is a vibrational tool widely used for the study not only of isolated and purified biomolecules but also of complex biological systems, including intact cells and tissues. One of the main advantages of this non-invasive approach is that it allows obtaining a molecular fingerprint of the sample under investigation in a rapid and label-free way. METHODS: Here we present an FTIR characterization of DRM fractions purified from the human breast cancer cells MCF-7, before and after treatment with the omega 3 fatty acid docosahexaenoic acid (DHA), which was found to promote membrane microdomain reorganization. RESULTS AND CONCLUSIONS: We will show that FTIR spectroscopy coupled with multivariate analysis enables to monitor changes in the composition of DRMs, induced in particular by the incorporation of DHA in cell membrane phospholipids. GENERAL SIGNIFICANCE: This study paves the way for a new label-free characterization of specific membrane domains within intact cells, which could provide complementary information to the fluorescence approaches presently used.
BACKGROUND: Detergent resistant membranes (DRMs) are a useful model system for the in vitro characterization of cell membrane domains. Indeed, DRMs provide a simple model to study the mechanisms underlying several key cell processes based on the interplay between specific cell membrane domains on one hand, and specific proteins and/or lipids on the other. Considering therefore their biological relevance, the development of methods capable to provide information on the composition and structure of membrane domains and to detect their modifications is highly desirable. In particular, Fourier transform infrared (FTIR) spectroscopy is a vibrational tool widely used for the study not only of isolated and purified biomolecules but also of complex biological systems, including intact cells and tissues. One of the main advantages of this non-invasive approach is that it allows obtaining a molecular fingerprint of the sample under investigation in a rapid and label-free way. METHODS: Here we present an FTIR characterization of DRM fractions purified from the humanbreast cancer cells MCF-7, before and after treatment with the omega 3 fatty aciddocosahexaenoic acid (DHA), which was found to promote membrane microdomain reorganization. RESULTS AND CONCLUSIONS: We will show that FTIR spectroscopy coupled with multivariate analysis enables to monitor changes in the composition of DRMs, induced in particular by the incorporation of DHA in cell membrane phospholipids. GENERAL SIGNIFICANCE: This study paves the way for a new label-free characterization of specific membrane domains within intact cells, which could provide complementary information to the fluorescence approaches presently used.
Authors: Marta J Fiołka; Paulina Czaplewska; Katarzyna Macur; Tomasz Buchwald; Jolanta Kutkowska; Roman Paduch; Zbigniew Kaczyński; Jerzy Wydrych; Teresa Urbanik-Sypniewska Journal: PLoS One Date: 2019-03-11 Impact factor: 3.240
Authors: R Molinaro; C Corbo; J O Martinez; F Taraballi; M Evangelopoulos; S Minardi; I K Yazdi; P Zhao; E De Rosa; M B Sherman; A De Vita; N E Toledano Furman; X Wang; A Parodi; E Tasciotti Journal: Nat Mater Date: 2016-05-23 Impact factor: 43.841