Chemical Penetration Enhancers Increase Hydrogen Peroxide Uptake in <i>C. elegans</i> for <i>In Vivo</i> Fast Photochemical Oxidation of Proteins

Espino, Jessica A.; Zhang, Zhihui; Jones, Lisa M.

doi:10.1021/acs.jproteome.0c00245

Cited by 18 publications

(35 citation statements)

References 41 publications

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“…Animals were grouped as mentioned in Table S2; AMY and AMYDOPA were applied on the epilated dermal side of animal skin by gentle rubbing and kept undisturbed. Skin irritancy caused by hydrolytic enzymes could be reduced by liposome coating, which prevents direct contact with skin . Infact, enhanced permeation of dextrans and insulin observed through the intercellular pathway and hair follicular route after trypsin treatment caused low skin irritation .…”

Section: Resultsmentioning

confidence: 99%

“…Skin irritancy caused by hydrolytic enzymes could be reduced by liposome coating, which prevents direct contact with skin. 56 Infact, enhanced permeation of dextrans and insulin observed through the intercellular pathway and hair follicular route after trypsin treatment caused low skin irritation. 57 However, in our study, there were no signs of allergic bleeding and lump formation of edema on the animal skin even after 24 h. We achieved maximum penetration of collagen within 15 min after treatment with 0.5 U of AMYDOPA.…”

Section: 3mentioning

confidence: 99%

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Enhanced Cellular Uptake and Sustained Transdermal Delivery of Collagen for Skin Regeneration

Pandurangan

Pooranachithra

Ramudu

et al. 2020

ACS Appl. Bio Mater.

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The present study reports a method for transporting high molecular weight collagen for skin regeneration. An independent engineered enzymatic vehicle that has the ability for efficient transdermal delivery of regenerative biomaterial was developed for tissue regeneration. Collagen has been well recognized as a skin regeneration molecule due to its interaction with the extracellular matrix to stimulate skin cell growth, proliferation, and differentiation. However, the transdermal delivery of collagen poses a significant challenge due to its high molecular weight as well as a lack of efficient approaches. Here, to improve the transdermal delivery efficiency, α-1,4-glycosidic hydrolase was engineered with genetically encoded 3,4-dihydroxy-L-phenylalanine, which enhanced its biological activity as revealed by microscale thermophoresis. The remodeled catalytic pocket resulted in enhanced substrate binding activity of the enzyme with a predominant glycosaminoglycan (chondroitin sulfate) present in the extracellular matrix of the skin. The engineered enzyme rapidly opened up the skin extracellular matrix fiber (15 min) to ferry collagen across the wall, without disturbing the cellular bundle architecture. Confocal microscopy indicated that macromolecules had diffused three times deeper into the engineered enzyme-treated skin than the native enzyme-treated skin. Gene expression, histopathology, and hematology analysis also supported the penetration of macromolecules. Cytotoxicity (mammalian cell culture) and in vivo (Caenorhabditis elegans and Rattus noryegicus) studies revealed that the congener enzyme could potentially be used as a penetration enhancer, which is of paramount importance for the multimillion cosmetic industries. Hence, it offers promise as a pharmaceutical enzyme for transdermal delivery bioenhancement and dermatological applications.

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Section: Resultsmentioning

confidence: 99%

Section: 3mentioning

confidence: 99%

Enhanced Cellular Uptake and Sustained Transdermal Delivery of Collagen for Skin Regeneration

Pandurangan

Pooranachithra

Ramudu

et al. 2020

ACS Appl. Bio Mater.

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“…This approach facilitates protein labelling within organelles and, in the case of C. elegans, within whole organisms, enabling structural interrogation under native conditions. [110][111][112]…”

Section: Hydroxyl Radical Footprinting-mass Spectrometry (Hrf-ms)mentioning

confidence: 99%

“…65,67,322 The FPOP derivative of HRF-MS has had similar in cell success over recent years, with a 2019 paper from Espino and Jones applying technique in vivo to determine protein solvent accessibility in C. elegans. [110][111][112] Given the prevalent use of C. elegans as a model organism, development of this workflow has exciting potential for the future study of disease mechanisms. The development of in vivo structural MS, however, is not limited to the peptide-based techniques, as developments have also been made in an attempt to bring protein-based methods into cells.…”

Section: Moving Structural Biology Towards In Cell Analysismentioning

confidence: 99%

Integration of Mass Spectrometry Data for Structural Biology

2021

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Mass spectrometry (MS) is increasingly being used to probe the structure and dynamics of proteins and the complexes they form with other macromolecules. There are now several specialised MS methods each with unique sample preparation, data acquisition and data processing protocols. Collectively these methods are referred to as structural MS and include, crosslinking-, hydrogen deuterium exchange-, hydroxyl radical footprinting-native, ion mobility-and top-down MS. Each of these provides a unique type of structural information, ranging from composition and stoichiometry through to residue level proximity and solvent accessibility. Structural MS has proved particularly beneficial in studying protein classes for which analysis by classic structural biology techniques proves challenging, such as glycosylated or intrinsically disordered proteins. To capture the structural details for a particular system, especially larger multiprotein complexes, more than one structural MS method with other structural and biophysical techniques is often required. Key to integrating these diverse data are computational strategies and software solutions to facilitate this process.We provide a background to the structural MS methods and briefly summarise other structural methods and how these are combined with MS. We then describe current state of the art approaches for the integration of structural MS data for structural biology. We quantify how often these methods are used together and provide examples where such combinations have been fruitful. To illustrate the power of integrative approaches, we discuss progress in solving the structures of the proteasome and the nuclear pore complex. We also discuss how information from structural MS, particularly pertaining to protein dynamics is not currently utilised in integrative workflows and how such information can provide a more accurate picture of the systems studied. We conclude by discussing new developments in the MS and computational fields that will further enable in-cell structural studies.

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“…Reagent access to the cells or organisms is one of the more challenging aspects of CL-MS experiments, yet it is essential to control as it largely determines the labeling efficiency, especially for the intracellular side of membrane proteins and for multicellular organisms. Jones and co-workers have found that the addition of CPEs can increase the uptake of labeling reagents via mild disruption to the lipid bilayer (Espino, Zhang, & Jones, 2020). These CPEs should not be toxic to the cell or organism within the working concentrations and ideally should not interfere with the labeling reagent.…”

Section: Chemical Penetration Enhancers (Cpes)mentioning

confidence: 99%

Membrane Protein Structures and Interactions From Covalent Labeling Coupled With Mass Spectrometry

Pan

Vachet

2020

Mass Spectrometry Reviews

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Membrane proteins are incredibly important biomolecules because they mediate interactions between a cell's external and internal environment. Obtaining information about membrane protein structure and interactions is thus important for understanding these essential biomolecules. Compared with the analyses of water-soluble proteins, the structural analysis of membrane proteins is more challenging owing to their unique chemical properties and the presence of lipid components that are necessary to solubilize them. The combination of covalent labeling (CL) and mass spectrometry (MS) has recently been applied with great success to study membrane protein structure and interactions. These studies have demonstrated the many advantages that CL-MS methods have over other traditional biophysical techniques. In this review, we discuss both amino acid-specific and non-specific labeling approaches and the special considerations needed to address the unique challenges associated with interrogating membrane proteins. This review highlights the aspects of this approach that require special care to be applied correctly and provides a comprehensive review of the membrane protein systems that have been studied by CL-MS.

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Chemical Penetration Enhancers Increase Hydrogen Peroxide Uptake in C. elegans for In Vivo Fast Photochemical Oxidation of Proteins

Cited by 18 publications

References 41 publications

Enhanced Cellular Uptake and Sustained Transdermal Delivery of Collagen for Skin Regeneration

Enhanced Cellular Uptake and Sustained Transdermal Delivery of Collagen for Skin Regeneration

Integration of Mass Spectrometry Data for Structural Biology

Membrane Protein Structures and Interactions From Covalent Labeling Coupled With Mass Spectrometry

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