Identification of protein secondary structures by laser induced autofluorescence: A study of urea and GnHCl induced protein denaturation

Siddaramaiah, Manjunath; Satyamoorthy, Kapaettu; Rao, Bola Sadashiva Satish; Roy, Suparna; Chandra, Subhash; Mahato, Krishna Kishore

doi:10.1016/j.saa.2016.11.017

Cited by 12 publications

(6 citation statements)

References 51 publications

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“…Excitation-dependent multicolor FL in the range of 450-550 nm was found in amyloid fibers and spherulites fabricated from β-LG from bovine milk and bovine insulin [96]. Recently, the use of laser-induced fibril FL properties as a characterization method for biological protein secondary structures, in addition to convention circular dichroism, was proposed [97]. Moreover, this FL effect allows the direct observation of the seeding, nucleation, and growth of amyloid fibrils at the earliest stages of this amyloidosis ( Figure 6) [85].…”

Section: Intrinsic Fluorescence In Amyloid Fibrilsmentioning

confidence: 99%

Fluorescence Phenomena in Amyloid and Amyloidogenic Bionanostructures

et al. 2020

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Nanoscale optical labeling is an advanced bioimaging tool. It is mostly based on fluorescence (FL) phenomena and enables the visualization of single biocells, bacteria, viruses, and biological tissues, providing monitoring of functional biosystems in vitro and in vivo, and the imaging-guided transportation of drug molecules. There is a variety of FL biolabels such as organic molecular dyes, genetically encoded fluorescent proteins (green fluorescent protein and homologs), semiconductor quantum dots, carbon dots, plasmonic metal gold-based nanostructures and more. In this review, a new generation of FL biolabels based on the recently found biophotonic effects of visible FL are described. This intrinsic FL phenomenon is observed in any peptide/protein materials folded into β-sheet secondary structures, irrespective of their composition, complexity, and origin. The FL effect has been observed both in natural amyloid fibrils, associated with neurodegenerative diseases (Alzheimer’s, Parkinson’s, and more), and diverse synthetic peptide/protein structures subjected to thermally induced biological refolding helix-like→β-sheet. This approach allowed us to develop a new generation of FL peptide/protein bionanodots radiating multicolor, tunable, visible FL, covering the entire visible spectrum in the range of 400–700 nm. Newly developed biocompatible nanoscale biomarkers are considered as a promising tool for emerging precise biomedicine and advanced medical nanotechnologies (high-resolution bioimaging, light diagnostics, therapy, optogenetics, and health monitoring).

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Section: Intrinsic Fluorescence In Amyloid Fibrilsmentioning

confidence: 99%

Fluorescence Phenomena in Amyloid and Amyloidogenic Bionanostructures

et al. 2020

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“…Besides, due to their complex structure, easy inactivation, and denaturation, it is difficult to modify additional active functions to them. [13,14] Therefore, based on the excellent biological properties of proteins, in order to enrich the functions of protein-based biomaterials, new methods to make specific functional modifications are expected, which enables the extension, the design and application of functional protein-based biomaterials.…”

mentioning

confidence: 99%

Injectable Polypeptide‐Protein Hydrogels for Promoting Infected Wound Healing

Cheng

Cai

et al. 2020

Adv Funct Materials

211

168

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Protein is the key composition of all tissues, which has also been widely used in tissue engineering due to its superior biocompatibility and low immunogenicity. However, natural protein usually lacks active functions such as vascularization, osteo‐induction, and neural differentiation, which limits its further applications as a functional biomaterial. Here, based on the mimetic extracellular matrix feature of bovine serum albumin, injectable polypeptide‐protein hydrogels with vascularization and antibacterial abilities are constructed successfully via coordinative cross‐linking of sulfydryl groups with silver ions (Ag+) for the regeneration of infected wound. In this protein hydrogel system, (Ag+), acting as crosslinkers, can not only provide a sterile microenvironment and a strong and robust antibacterial ability but also introduce K2(SL)6K2 (KK) polypeptide, which endows the hydrogel with vascularization behavior. Furthermore, the in vivo data show that the polypeptide‐protein hydrogel has a considerable collagen deposition and vascularization abilities in the early stage of wound healing, resulting in rapid new tissue regeneration featured with newly appeared hair follicles. Altogether, this newly developed multifunctional 3D polypeptide‐protein hydrogel with vascularization, antibacterial properties, and hair follicle promotion can be a promising approach in biomedical fields such as infected wound healing.

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“…The spectral unmixing process benefits from the acquisition of autofluorescence spectra, characterized by the separation into autofluorescence components and optimization of each component’s spectrum from actual sample data. The autofluorescence spectra from different cells vary based on the contained autofluorescence molecules 22 and are also influenced by tissue fixation conditions 23, 24 . The separation of autofluorescence spectrum components aligns with cellular type differences and is optimized from spectral data acquired from unstained slides, starting with five fluorescence spectra: NADH and FAD as fluorescent nucleotides, arachidonic acid for lipids, porphyrins constituting hemoglobin and cytochrome, and the mounting agent (Fig.…”

Section: Resultsmentioning

confidence: 99%

Spatial Clustering Analysis with Spectral Imaging-based Single-Step Multiplex Immunofluorescence (SISS-mIF)

Nakamura,

Kaneko,

Taguchi

et al. 2024

Preprint

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Precision medicine, anchored in spatial biology, is essential for the accurate diagnosis of cancer and prediction of drug responses. We have introduced the Spectral Imaging-based Single-Step Multiplex Immunofluorescence (SISS-mIF) technique, which leverages hyperspectral imaging to simultaneously capture fluorescence spectra. This approach automatically optimizes tissue autofluorescence spectra for each image, facilitating the use of fluorescent direct-labeled antibodies for multicolor staining in a single step. Unlike conventional methods, images are outputted as antibody counts rather than fluorescence intensity, allowing for consistent comparisons under different imaging conditions. We demonstrate that this technique allows for identical cell detection of CD3, CD5, and CD7 in T-cell lymphoma on a single slide. The utilization of fluorescent direct-labeled antibodies enables the triple staining of CD3, CD5, and CD7 without cross-reactivity, maintaining the same intensity as single stains. Moreover, we developed a joint Non-Negative Matrix Factorization-based Spatial Clustering Analysis (jNMF-SCA) with a modified spectral unmixing system, highlighting its potential as a supportive diagnostic tool for T-cell lymphoma.

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Identification of protein secondary structures by laser induced autofluorescence: A study of urea and GnHCl induced protein denaturation

Cited by 12 publications

References 51 publications

Fluorescence Phenomena in Amyloid and Amyloidogenic Bionanostructures

Fluorescence Phenomena in Amyloid and Amyloidogenic Bionanostructures

Injectable Polypeptide‐Protein Hydrogels for Promoting Infected Wound Healing

Spatial Clustering Analysis with Spectral Imaging-based Single-Step Multiplex Immunofluorescence (SISS-mIF)

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