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      Nanoscale Infrared Spectroscopy and Chemometrics Enable Detection of Intracellular Protein Distribution

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          Abstract

          Determination of the intracellular location of proteins is one of the fundamental tasks of microbiology. Conventionally, label-based microscopy and super-resolution techniques are employed. In this work, we demonstrate a new technique that can determine intracellular protein distribution at nanometer spatial resolution. This method combines nanoscale spatial resolution chemical imaging using the photothermal-induced resonance (PTIR) technique with multivariate modeling to reveal the intracellular distribution of cell components. Here, we demonstrate its viability by imaging the distribution of major cellulases and xylanases in Trichoderma reesei using the colocation of a fluorescent label (enhanced yellow fluorescence protein, EYFP) with the target enzymes to calibrate the chemometric model. The obtained partial least squares model successfully shows the distribution of these proteins inside the cell and opens the door for further studies on protein secretion mechanisms using PTIR.

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          Most cited references78

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          Fourier transform infrared spectroscopic analysis of protein secondary structures.

          Infrared spectroscopy is one of the oldest and well established experimental techniques for the analysis of secondary structure of polypeptides and proteins. It is convenient, non-destructive, requires less sample preparation, and can be used under a wide variety of conditions. This review introduces the recent developments in Fourier transform infrared (FTIR) spectroscopy technique and its applications to protein structural studies. The experimental skills, data analysis, and correlations between the FTIR spectroscopic bands and protein secondary structure components are discussed. The applications of FTIR to the secondary structure analysis, conformational changes, structural dynamics and stability studies of proteins are also discussed.
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            Breaking the diffraction barrier: super-resolution imaging of cells.

            Anyone who has used a light microscope has wished that its resolution could be a little better. Now, after centuries of gradual improvements, fluorescence microscopy has made a quantum leap in its resolving power due, in large part, to advancements over the past several years in a new area of research called super-resolution fluorescence microscopy. In this Primer, we explain the principles of various super-resolution approaches, such as STED, (S)SIM, and STORM/(F)PALM. Then, we describe recent applications of super-resolution microscopy in cells, which demonstrate how these approaches are beginning to provide new insights into cell biology, microbiology, and neurobiology. Copyright © 2010 Elsevier Inc. All rights reserved.
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              Infrared spectroscopy of proteins.

              This review discusses the application of infrared spectroscopy to the study of proteins. The focus is on the mid-infrared spectral region and the study of protein reactions by reaction-induced infrared difference spectroscopy.
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                Author and article information

                Journal
                Anal Chem
                Anal Chem
                ac
                ancham
                Analytical Chemistry
                American Chemical Society
                0003-2700
                1520-6882
                01 December 2020
                15 December 2020
                : 92
                : 24
                : 15719-15725
                Affiliations
                []Institute of Chemical Technologies and Analytics, TU Wien , Vienna 1060, Austria
                []Institute of Chemical, Environmental and Bioscience Engineering, TU Wien , Vienna 1060, Austria
                Author notes
                Article
                10.1021/acs.analchem.0c02228
                7745202
                33259186
                5ad8a6a9-f57d-40b6-a84d-a13d8af4479f
                © 2020 American Chemical Society

                This is an open access article published under a Creative Commons Attribution (CC-BY) License, which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.

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                Custom metadata
                ac0c02228
                ac0c02228

                Analytical chemistry
                Analytical chemistry

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