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Comparative Characterization of Plasmodium falciparum Hsp70-1 Relative to E. coli DnaK Reveals the Functional Specificity of the Parasite Chaperone

Publication year: 2020
Authors: Lebepe C. 1, Matambanadzo P. 1, Makhoba X. 2, Achilonu I. 3, Zininga T. 1,4, Shonhai A. 1,*
Published in: Biomolecules 2020, Vol. 10(6), p. 856
Hsp70 is a conserved molecular chaperone. How Hsp70 exhibits specialized functions across species remains to be understood. Plasmodium falciparum Hsp70-1 (PfHsp70-1) and Escherichia coli DnaK are cytosol localized molecular chaperones that are important for the survival of these two organisms. [...]

PEDOT-polyamine composite films for bioelectrochemical platforms -flexible and easy to derivatize

Publication year: 2020
Authors: Sappia L. +a, Piccinini E. +a, von Binderling C. a, Knoll W. bcd, Marmisollé W. *a, Azzorini O. *acd
Published in: Material Sciece and Engineering: C, 2020, Vol. 109, p.110575
We report a straightforward route for the preparation of flexible, electrochemically stable and easily functionalizable poly(3,4-ethylenedioxythiophene) (PEDOT) composite films deposited on PET foils as biosensing platforms. For this purpose, poly(allylamine) hydrochloride (PAH) was blended with PEDOT to provide amine-bearing sites for further biofunctionalization as well as to improve the mechanical properties of the films.[...]

It takes two to tango - The case of thebaine 6-O-demethylase

Publication year: 2020
Authors: Kachhap S. a*, Wojdyla Z. a*, Komorek P. a, Kluza A. a, Kurpiewska K. ab, Jachimska B. a,Borowski T.a
Published in: nternational Journal of Biological Macromolecules, 2020, Vol. 163, p. 718-729
Thebaine 6-O-demethylase (T6ODM) is an Fe(II)/2-oxoglutarate-dependent dioxygenase catalysing two oxidative O-demethylation reactions in morphine biosynthesis. Its crystal structure revealed a large active site pocket which is at least two times larger than necessary to accommodate a substrate (thebaine or oripavine) molecule.[...]

Surface Preconditioning Influences the Antifouling Capabilities of Zwitterionic and Nonionic Polymer Brushes

Publication year: 2020
Authors: Víšová I. 1, Vrabcová M. 1, Forinová M. 1, Zhigunová Y. 1, Mironov V. 1, Houska . 1, Bittrich E. 2, Eichhorn K. 2, Hashim H. 3,4, Schovánek P. 1,5, Dejneka A. 1, Vaisocherová - Lísalová H. 1
Published in: Langmuir, 2020, Vol. 36, 29, 8485–8493
Polymer brushes not only represent emerging surface platforms for numerous bioanalytical and biological applications but also create advanced surface-tethered systems to mimic real-life biological processes. In particular, zwitterionic and nonionic polymer brushes have been intensively studied because of their extraordinary resistance to nonspecific adsorption of biomolecules (antifouling characteristics) as well as the ability to be functionalized with bioactive molecules.[...]

Label-Free Analysis with Multiple Parameters Separates G Protein-Coupled Receptor Signaling Pathways

Publication year: 2020
Authors: Temu Suutari*, Sabrina N. Rahman, Henry F. Vischer, Dick van Iperen, Arto Merivaara, Marjo Yliperttula, Rob Leurs, Jeroen Kool, Tapani Viitala*
Published in: Analytical Chemistry, 2020, Vol. 92, 21, p. 14509–14516
Real-time label-free techniques are used to profile G protein-coupled receptor (GPCR) signaling pathways in living cells. However, interpreting the label-free signal responses is challenging, [...]

Modulation of Living Cell Behavior with Ultra‐Low Fouling Polymer Brush Interfaces

Publication year: 2020
Authors: Víšová I., Smolková B., Uzhytchak M., Vrabcová M., Zhigunova Y., Houska M. ,Surman F., de los Santos Pereira A.,Lunov O., Dejneka A., Vaisocherová‐Lísalová H.
Published in: Macromolecular Bioscience, 2020, Vol. 20, 3, p. 1900351
Ultra‐low fouling and functionalizable coatings represent emerging surface platforms for various analytical and biomedical applications such as those involving examination of cellular interactions in their native environments. Ultra‐low fouling surface platforms as advanced interfaces enabling modulation of behavior of living cells via tuning surface physicochemical properties are presented and studied.[...]

Asymmetric flow field-flow fractionation coupled to surface plasmon resonance detection for analysis of therapeutic proteins in blood serum

Publication year: 2020
Authors: Mats Leeman 1, Willem M. Albers 2, Radoslaw Bombera 2, Johana Kuncova-Kallio 2, Jussipekka Tuppurainen 2, Lars Nilsson 3
Published in: Analytical and Bioanalytical Chemistry, 2020
Coupling of surface plasmon resonance (SPR) detection to asymmetric flow field-flow fractionation (AF4) offers the possibility to study active fractions of bio-separations on real samples, such as [...]

Generic High-Capacity Electrochemical Capture and Release of Proteins by Polyelectrolyte Brushes

Publication year: 2020
Authors: Ferrand-Drake del Castillo G., Adali Z., L. N. Hailes R., Xiong K., Dahlin A.*
Published in: Preprint in Chem RXIV, 2020
We present a polyelectrolyte brush electrode with high-capacity for protein immobilization that captures and releases proteins by an electrochemical potential. Central to our concept is the use of polyelectrolyte brushes that switch reversibly by electrochemistry.[...]

Functional Plasma Polymerized Surfaces for Biosensing

Publication year: 2020
Authors: Makhneva* E., Barillas L 1., Farka Z. 2, Pastucha M. 2, Skládal P. 2, Weltmann K.D. 1, Fricke K. 1
Published in: Applied Material Interfaces, 2020, Vol.12, 14, p.17100–17112
The capabilities of biosensors for fast, economic, and user-friendly analysis of complex samples has led to the exploitation of analytical devices for detection, quantification, and monitoring of specific chemical species for various applications. For a sufficiently high surface reactivity toward the adopted bioreceptors, a thin functional layer is required to enable coupling of the target biomolecules and to provide good stability in the presence of a sample matrix.[...]

Matter‐tag: A universal immobilization platform for enzymes on polymers, metals, and silicon‐based materials

Publication year: 2020
Authors: Dedisch S. 12, Wiens A. 2, Davari M.D. 2, Söder D. 1, Rodriguez‐Emmenegger C. 13, Jakob F. 12, Schwaneberg U.*12
Published in: Biotechnology and Bioengineering, 2020, Vol. 117, Issue1, p. 49-61
Enzyme immobilization is extensively studied to improve enzyme properties in catalysis and analytical applications. Here, we introduce a simple and versatile enzyme immobilization platform based on adhesion‐promoting peptides, namely Matter‐tags. Matter‐tags immobilize enzymes in an oriented way as a dense monolayer.[...]

Enhancing the Sensitivity of Biotinylated Surfaces by Tailoring the Design of the Mixed Self-Assembled Monolayer Synthesis

Publication year: 2020
Authors: Blasi D. ‡†, Sarcina L. ‡§, Tricase A. ‡§, Stefanachi A. ‖, Leonetti F.*‖, Alberga D. $, Mangiatordi* Ͱ, Manoli K. †§, Scamarcio G. ◊∫, Picca R.A. *†§, Torsi L. †§#
Published in: ACS Omega 2020, Vol. 5, 27, p. 16762–16771
Thiolated self-assembled monolayers (SAMs) are typically used to anchor on a gold surface biomolecules serving as recognition elements for biosensor applications. Here, the design and synthesis of N-(2-hydroxyethyl)-3-mercaptopropanamide (NMPA) in biotinylated mixed SAMs is proposed as an alternative strategy with respect to on-site multistep functionalization of SAMs prepared from solutions of commercially available thiols.[...]

In situ analysis of liposome hard and soft protein corona structure and composition in a single label-free workflow

Publication year: 2020
Authors: Kari O. *ab, Ndika J. c, Parkkila P. b, Louna A. a, Lajunen T. ǂa, Puustinen A. d, Viitala T. *§b, Alenius H. ce, Urtti A. af
Published in: Nanoscale, 2020, Vol. 12, p. 1728
Methodological constraints have limited our ability to study protein corona formation, slowing nanomedicine development and their successful translation into the clinic. We determined hard and soft corona structural properties along with the corresponding proteomic compositions on liposomes in a label-free workflow: surface plasmon resonance and a custom biosensor for in situ structure determination on liposomes and corona separation, and proteomics using sensitive nanoliquid chromatography tandem mass spectrometry with open-source bioinformatics platforms.[...]

Nickel (II) phthalocyanine-tetrasulfonic-Au nanoparticles nanocomposite film for tartrazine electrochemical sensing

Publication year: 2020
Authors: de Lima L. 1, Maciel C. 2, Ferreira A. 1, de Almeida J. 2, Ferreira M. 1*
Published in: Materials Letters, 2020 Vol. 262, p. 127186
In this work, we report the analytical detection of tartrazine through differential pulse voltammetry (DPV) using gold nanoparticles (AuNPs) and Nickel(II)-phthalocyanine-tetrasulfonic (NiTsPc). The Layer-by-Layer (LbL) film was constructed following the Au-(AuNPs-PAH/NiTsPc)5 architecture.[...]

Surfactant Interactions with Protein-Coated Surfaces: Comparison between Colloidal and Macroscopically Flat Surfaces

Publication year: 2020
Authors: Mateos H. 1,*, Valentini A. 2, Lopez F. 3, Palazzo G. 4
Published in: Biomimetics 2020, Vol. 5(3), p. 31
Surface interactions with polymers or proteins are extensively studied in a range of industrial and biomedical applications to control surface modification, cleaning, or biofilm formation. In this study we compare surfactant interactions with protein-coated silica surfaces differing in the degree of curvature (macroscopically flat and colloidal nanometric spheres). The interaction with a flat surface was probed by means of surface plasmon resonance (SPR) while dynamic light scattering (DLS) was used to study the interaction with colloidal SiO2 (radius 15 nm).Surface interactions with polymers or proteins are extensively studied in a range of industrial and biomedical applications to control surface modification, cleaning, or biofilm formation. In this study we compare surfactant interactions with protein-coated silica surfaces differing in the degree of curvature (macroscopically flat and colloidal nanometric spheres). The interaction with a flat surface was probed by means of surface plasmon resonance (SPR) while dynamic light scattering (DLS) was used to study the interaction with colloidal SiO2 (radius 15 nm).[...]

Layer-by-Layer nanostructured films of magnetite nanoparticles and polypyrrole towards synergistic effect on methylparaben electrochemical detection

Publication year: 2020
Authors: de Lima L., Daikuzono C., Miyazaki C., Pereira E., Ferreira M.*
Published in: Applied Surface Science 2020, Vol. 505, p. 144278
The sustained use of cosmetics, hygiene products and medicines containing methylparaben (MeP) can be related to cases of cancer in women and children, which is why it is important to develop simple ways to monitor this compound. Gold electrodes were modified with Layer-by-Layer (LbL) films of magnetite nanoparticles (MNP) and polypyrrole (Ppy) for electrochemical determination of MeP.[...]

Delineating the Molecular Basis of the Calmodulin–bMunc13-2 Interaction by Cross-Linking/Mass Spectrometry—Evidence for a Novel CaM Binding Motif in bMunc13-2

Publication year: 2020
Authors: Piotrowski C. 1, Moretti R. 2, Ihling C. 1,Haedicke A. 3†, Liepold T. 4, Lipstein N. 5, Meiler J. 2, Jahn O. 4,*, Sinz A. 1,*
Published in: Cells, 2020, Vol.9 (1) , p.136
Exploring the interactions between the Ca2+ binding protein calmodulin (CaM) and its target proteins remains a challenging task. Members of the Munc13 protein family play an essential role in short-term synaptic plasticity, modulated via the interaction with CaM at the presynaptic compartment. [...]

An investigation of the synergistic effect between magnetite nanoparticles and polypyrrole in nanostructured layer‐by‐layer films

Publication year: 2020
Authors: de Lima L. 1,2, Maciel C. 3, Ferreira A. 1, Rubira R. 4, Constantino C. 4, Ferreira M. *1 1M.
Published in: Journal of Applied Polymer Science, 2020
In this paper, we report the controlled fabrication of layer‐by‐layer (LbL) films deposited on gold substrates with three different supramolecular architectures using polypyrrole (Ppy) and magnetite nanoparticles (Fe3O4‐np), besides conventional poly(allylamine hydrochloride) (PAH) e poly(vinyl sulfonic acid) (PVS) polyelectrolytes, demonstrating the synergistic effect between Ppy and Fe3O4‐np such as a result of their interaction.[...]

Single-vesicle imaging reveals lipid-selective and stepwise membrane disruption by monomeric α-synuclein

Publication year: 2020
Authors: Hannestad J. a, Rocha S. b, Agnarsson B. a, Zhdanov V. ac, Wittung-Stafshede P. *b, Höök F. *a
Published in: Proceedings of National Academy of Sciences PNAS, 2020, Vol. 117, 25, p.14178-14186
The interaction of the neuronal protein α-synuclein with lipid membranes appears crucial in the context of Parkinson’s disease, but the underlying mechanistic details, including the roles of different lipids in pathogenic protein aggregation and membrane disruption, remain elusive. Here, we used single-vesicle resolution fluorescence and label-free scattering microscopy to investigate the interaction kinetics of monomeric α-synuclein with surface-tethered vesicles composed of different negatively charged lipids.[...]

Ultra-low HIV-1 p24 detection limits with a bioelectronic sensor

Publication year: 2020
Authors: Macchia E. 12, Sarcina L. 1, Picca R.A. 1, Manoli K. 1, Di Franco C. 1, Scamarcio G. 34 Torsi L. 125
Published in: Analytical and Bioanalytical Chemistry, 2020, Vol. 412, p. 811–818
Early diagnosis of the infection caused by human immunodeficiency virus type-1 (HIV-1) is vital to achieve efficient therapeutic treatment and limit the disease spreading when the viremia is at its highest level. To this end, a point-of-care HIV-1 detection carried out with label-free, low-cost, and ultra-sensitive screening technologies would be of great relevance.[...]

Label-free characterization and real-time monitoring of cell uptake of extracellular vesicles

Publication year: 2020
Authors: Koponen A.ab, Kerkelä E. a, Rojalin T. b, Lázaro-Ibáñez E. b, Suutari T. b, Saari H.b, Siljander P.c, Yliperttula M. b, Laitinen S. a, Viitala T. b
Published in: Biosensors and Bioelectronics, 2020, Vol. 168, p. 112510
Extracellular vesicles (EVs) have the ability to function as molecular vehicles and could therefore be harnessed to deliver drugs to target cells in diseases such as cancer. The composition of EVs determines their function as well as their interactions with cells, which consequently affects the cell uptake efficacy of EVs.[...]