Raman spectroscopy characterization of the major classes of plasma lipoproteins

Ricciardi, Alessandra; Piuri, Gabriele; Porta, Matteo Della; Mazzucchelli, Serena; Bonizzi, Arianna; Truffi, Marta; Sevieri, Marta; Allevi, Raffaele; Corsi, Fabio; Cazzola, Roberta; Morasso, Carlo

doi:10.1016/j.vibspec.2020.103073

Cited by 20 publications

(30 citation statements)

References 31 publications

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“…Raman spectroscopy has been previously used to sensitively distinguish lipoprotein variants. 55 Here, samples are compared throughout using principal component analysis (PCA), which we have previously reported for reducing dimensionality of Raman spectral datasets. 56,57 PCA generates an intuitive visualization of multivariate data, capturing as much variability as possible and conserving the pertinent information responsible for the major sources of data variability.…”

Section: Resultsmentioning

confidence: 99%

Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination

et al. 2021

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Section: Resultsmentioning

confidence: 99%

Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination

et al. 2021

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“…Recently, modern Raman instruments coupled with microscopy and multivariate statistical analysis have made Raman spectroscopy a viable approach for the fast characterization of lipoproteins [54].…”

Section: Methods Of Hdl Measurement In the Laboratorymentioning

confidence: 99%

“…As is currently the use of cholesterol fractions, the determination of TG concentration into the different classes of lipoproteins could provide easy-to-interpret clinical indications for identifying dysfunctional HDL [21,56]. Similarly, Raman spectroscopy could be a quick and advantageous approach for the characterization of lipoproteins that allows obtaining simultaneous information about the biomolecules composing lipoproteins: cholesterol, CE, TG, membrane lipids, unsaturated fatty acid, carotenoids, proteins, and their relative amount without any sample preparation [54]. In addition, Raman spectroscopy allows the determination of the ratio between TGs and CEs and the unsaturation degree of surface lipid fatty acids, two factors that influence the degree of fluidity of the hydrophobic core and hydrophilic surface envelope of HDLs, respectively.…”

Section: Methods Of Hdl Measurement In the Laboratorymentioning

confidence: 99%

HDL Dysfunctionality: Clinical Relevance of Quality Rather Than Quantity

et al. 2021

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High-density lipoproteins (HDLs) represent a class of lipoproteins very heterogeneous in structure, composition, and biological functions, which carry out reverse cholesterol transport, antioxidant, anti-inflammatory, antithrombotic, and vasodilator actions. Despite the evidence suggesting a clear inverse relationship between HDL cholesterol (HDL-c) concentration and the risk for cardiovascular disease, plasma HDL cholesterol levels do not predict the functionality and composition of HDLs. The importance of defining both the amount of cholesterol transported and lipoprotein functionality has recently been highlighted. Indeed, different clinical conditions such as obesity, diabetes mellitus type 2 (T2DM), and cardiovascular disease (CVD) can alter the HDL functionality, converting normal HDLs into dysfunctional ones, undergoing structural changes, and exhibiting proinflammatory, pro-oxidant, prothrombotic, and proapoptotic properties. The aim of the current review is to summarize the actual knowledge concerning the physical–chemical alteration of HDLs related to their functions, which have been found to be relevant in several pathological conditions associated with systemic inflammation and oxidative stress.

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“…b. Curve-fitting procedure: Curve fitting of the Amide I band and assignment to the structural components were performed essentially as described by Maiti et al, (2003) 22 . Curve fitting was performed via Origin (version 2022) 20 . c. Principle component analysis (PCA): PCA was applied on Raman spectroscopy data with 10 spectra, being satisfactorily described by a principle component number 21.…”

Section: Raman Spectroscopymentioning

confidence: 99%

Biophysical Characterization of an Essential Mammalian Protein; Transcription Termination Factor I (TTF1)

Tiwari

Singh

2022

Preprint

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Mammalian Transcription Terminator Factor 1 (TTF1) is an essential protein which plays diverse cellular physiological functions like transcription regulation (both initiation and termination), replication fork blockage, chromatin remodelling, DNA damage repair etc. Hence, understanding the structure and mechanism conferred by its variable confirmations becomes significantly important. But so far, almost nothing is known about the structure of either the full length protein or any of its domain in isolation. Moving towards achieving the above goals, our lab has codon optimised, expressed and purified N-terminal 190 amino acid deleted TTF1 (∆N190TTF1) protein, since full length protein even after multiple trials could not be purified in soluble form. In this article, we have characterized this essential protein by studying its homogeneity, molecular size and secondary structure using tools like dynamic light scattering (DLS), circular dichroism (CD) spectroscopy, Raman spectroscopy and atomic force microscopy (AFM). By CD and DLS we have shown that the purified protein is homogenous and soluble. CD spectroscopy also revealed that ∆N190TTF1 is a helical protein which was further confirmed by analysis of Raman spectra and Amide I region deconvolution studies. AFM imaging data discovered the size of single protein molecule to be 94 nm which is in agreement with the size determined by the DLS study as well. Our structural and biophysical characterization of this essential protein will open avenues towards solving the structure to atomic resolution and also will encourage the research to investigate the mechanism behind its diverse functions attributed to its various domains.

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Raman spectroscopy characterization of the major classes of plasma lipoproteins

Cited by 20 publications

References 31 publications

Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination

Surface enhanced Raman scattering of extracellular vesicles for cancer diagnostics despite isolation dependent lipoprotein contamination

HDL Dysfunctionality: Clinical Relevance of Quality Rather Than Quantity

Biophysical Characterization of an Essential Mammalian Protein; Transcription Termination Factor I (TTF1)

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