Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 17 de 17
Filter
Add more filters










Publication year range
1.
Nanotechnology ; 34(23)2023 Mar 20.
Article in English | MEDLINE | ID: mdl-36863009

ABSTRACT

Following the demonstration of laser-induced ultrafast demagnetization in ferromagnetic nickel, several theoretical and phenomenological propositions have sought to uncover its underlying physics. In this work we revisit the three temperature model (3TM) and the microscopic three temperature model (M3TM) to perform a comparative analysis of ultrafast demagnetization in 20 nm thick cobalt, nickel and permalloy thin films measured using an all-optical pump-probe technique. In addition to the ultrafast dynamics at the femtosecond timescales, the nanosecond magnetization precession and damping are recorded at various pump excitation fluences revealing a fluence-dependent enhancement in both the demagnetization times and the damping factors. We confirm that the Curie temperature to magnetic moment ratio of a given system acts as a figure of merit for the demagnetization time, while the demagnetization times and damping factors show an apparent sensitivity to the density of states at the Fermi level for a given system. Further, from numerical simulations of the ultrafast demagnetization based on both the 3TM and the M3TM, we extract the reservoir coupling parameters that best reproduce the experimental data and estimate the value of the spin flip scattering probability for each system. We discuss how the fluence-dependence of inter-reservoir coupling parameters so extracted may reflect a role played by nonthermal electrons in the magnetization dynamics at low laser fluences.

2.
Luminescence ; 38(7): 845-866, 2023 Jul.
Article in English | MEDLINE | ID: mdl-35419945

ABSTRACT

Carbon quantum dots (CQDs) are promising carbonaceous nanomaterials fortuitously discovered in 2004. CQDs are the rising stars in the nanotechnology ensemble because of their unique properties and widespread applications in sensing, imaging, medicine, catalysis, and optoelectronics. CQDs are notable for their excellent solubility and effective luminescence and, as a result, they are also known as carbon nanolights. Many strategies are used for the efficient and economical preparation of CQDs; however, CQDs prepared from waste or green sustainable methods have greater requirements due to their safety and ease of synthesis. Sustainable chemical strategies for CQDs have been developed, emphasizing green synthetic methodologies based on 'top-down' and 'bottom-up' approaches. This review summarizes many such studies relevant to the development of sustainable methods for photoluminescent CQDs. Furthermore, we have emphasized recent advances in CQDs' photoluminescence applications in chemical and biological fields. Finally, a brief overview of synthetic processes using the green source and their associated applications are tabulated, providing a clear understanding of the new optoelectronic materials.


Subject(s)
Quantum Dots , Quantum Dots/chemistry , Carbon/chemistry , Luminescence , Catalysis
3.
Nanotechnology ; 34(13)2023 Jan 20.
Article in English | MEDLINE | ID: mdl-36571848

ABSTRACT

Reconfigurable magnonics have attracted intense interest due to their myriad advantages including energy efficiency, easy tunability and miniaturization of on-chip data communication and processing devices. Here, we demonstrate efficient reconfigurability of spin-wave (SW) dynamics as well as SW avoided crossing by varying bias magnetic field orientation in triangular shaped Ni80Fe20nanodot arrays. In particular, for a range of in-plane angles of bias field, we achieve mutual coherence between two lower frequency modes leading to a drastic modification in the ferromagnetic resonance frequency. Significant modification in magnetic stray field distribution is observed at the avoided crossing regime due to anisotropic dipolar interaction between two neighbouring dots. Furthermore, using micromagnetic simulations we demonstrate that the hybrid SW modes propagate longer through an array as opposed to the non-interacting modes present in this system, indicating the possibility of coherent energy transfer of hybrid magnon modes. This result paves the way for the development of integrated on-chip magnonic devices operating in the gigahertz frequency regime.

4.
3 Biotech ; 12(4): 92, 2022 Apr.
Article in English | MEDLINE | ID: mdl-35342680

ABSTRACT

Electrospinning is an electrostatic fiber fabrication technique that operates by the application of a strong electric field on polymer solution or melts. It is used to fabricate fibers whose size lies in the range of few microns to the nanometer range. Historic development of electrospinning has evinced attention due to its outstanding attributes such as small diameter, excellent pore inter-connectivity, high porosity, and high surface-to-volume ratio. This review aims to highlight the theory behind electrospinning and the machine setup with a detailed discussion about the processing parameters. It discusses the latest innovations in natural protein-based electrospun nanofibers for health care applications. Various plant- and animal-based proteins have been discussed with detailed sample preparation and corresponding processing parameters. The usage of these electrospun nanofibers in regenerative medicine and drug delivery has also been discussed. Some technical innovations in electrospinning techniques such as emulsion electrospinning and coaxial electrospinning have been highlighted. Coaxial electrospun core-shell nanofibers have the potential to be utilized as an advanced nano-architecture for sustained release targeted delivery as well as for regenerative medicine. Healthcare applications of nanofibers formed via emulsion and coaxial electrospinning have been discussed briefly. Electrospun nanofibers have still much scope for commercialization on large scale. Some of the available wound-dressing materials have been discussed in brief.

5.
Adv Sci (Weinh) ; 9(8): e2104644, 2022 Mar.
Article in English | MEDLINE | ID: mdl-35043603

ABSTRACT

Tripartite coupling between phonons, magnons, and photons in a periodic array of elliptical magnetostrictive nanomagnets delineated on a piezoelectric substrate to form a 2D two-phase multiferroic crystal is investigated. Surface acoustic waves (SAW) (phonons) of 5-35 GHz frequency launched into the substrate cause the magnetizations of the nanomagnets to precess at the frequency of the wave, giving rise to confined spin-wave modes (magnons) within the nanomagnets. The spin waves, in turn, radiate electromagnetic waves (photons) into the surrounding space at the SAW frequency. Here, the phonons couple into magnons, which then couple into photons. This tripartite phonon-magnon-photon coupling is thus exploited to implement an extreme sub-wavelength electromagnetic antenna whose measured radiation efficiency and antenna gain exceed the approximate theoretical limits for traditional antennas of the same dimensions by more than two orders of magnitude at some frequencies. Micro-magnetic simulations are in excellent agreement with experimental observations and provide insight into the spin-wave modes that couple into radiating electromagnetic modes to implement the antenna.

6.
Nanoscale ; 13(32): 13709-13718, 2021 Aug 28.
Article in English | MEDLINE | ID: mdl-34477646

ABSTRACT

Graphene/ferromagnet hybrid heterostructures are important building blocks of spintronics due to the unique ability of graphene to transport spin current over unprecedented distances and possible increase in its spin-orbit coupling due to proximity and hybridization. Here, we present magnetization dynamics over a femtosecond to nanosecond timescale by employing an all-optical time-resolved magneto-optical Kerr effect technique in single-layer graphene (SLG)/CoFeB thin films with varying CoFeB thickness and compared them with reference CoFeB thin films without an SLG underlayer. Gilbert damping variation with CoFeB thickness is modelled to extract spin-mixing conductance for the SLG/CoFeB interface and isolate the two-magnon scattering contribution from spin pumping. In SLG/CoFeB, we have established an inverse relationship between ultrafast demagnetization time (τm) and the Gilbert damping parameter (α) induced by interfacial spin accumulation and pure spin-current transport via a spin pumping mechanism. This systematic study of ultrafast demagnetization in SLG/CoFeB heterostructures and its connection with magnetic damping can help to design graphene-based ultrahigh-speed spintronic devices.

7.
Nanoscale ; 13(22): 10016-10023, 2021 Jun 14.
Article in English | MEDLINE | ID: mdl-34037043

ABSTRACT

Using time-resolved magneto-optical Kerr effect (TR-MOKE) microscopy, we demonstrate surface-acoustic-wave (SAW) induced resonant amplification of intrinsic spin-wave (SW) modes, as well as generation of new extrinsic or driven modes at the SAW frequency, in a densely packed two-dimensional array of elliptical Co nanomagnets fabricated on a piezoelectric LiNbO3 substrate. This system can efficiently serve as a magnonic crystal (MC), where the intrinsic shape anisotropy and the strong inter-element magnetostatic interaction trigger the incoherent precession of the nanomagnets' magnetization in the absence of any bias magnetic field, giving rise to the 'intrinsic' SW modes. The magnetoelastic coupling leads to a rich variety of SW phenomena when the SAW is launched along the major axis of the nanomagnets, such as 4-7 times amplification of intrinsic modes (at 3, 4, 7 and 10 GHz) when the applied SAW frequencies are resonant with these frequencies, and the generation of new extrinsic modes at non-resonant SAW frequencies. However, when the SAW is launched along the minor axis, a dominant driven mode appears at the applied SAW frequency. This reveals that the magnetoelastic coupling between SW and SAW is anisotropic in nature. Micromagnetic simulation results are in qualitative agreement with the experimental observations and elucidate the underlying dynamics. Our findings lay the groundwork for bias-field free magnonics, where the SW behavior is efficiently tuned by SAWs. It has important applications in the design of energy efficient on-chip microwave devices, SW logic, and extreme sub-wavelength ultra-miniaturized microwave antennas for embedded applications.

8.
ACS Appl Mater Interfaces ; 13(17): 20875-20884, 2021 May 05.
Article in English | MEDLINE | ID: mdl-33886256

ABSTRACT

Pure spin current has transformed the research field of conventional spintronics due to its various advantages, including energy efficiency. An efficient mechanism for generation of pure spin current is spin pumping, and high effective spin-mixing conductance (Geff) and interfacial spin transparency (T) are essential for its higher efficiency. By employing the time-resolved magneto-optical Kerr effect technique, we report here a giant value of T in substrate/W (t)/Co20Fe60B20 (d)/SiO2 (2 nm) thin-film heterostructures in the beta-tungsten (ß-W) phase. We extract the spin diffusion length of W and spin-mixing conductance of the W/CoFeB interface from the variation of damping as a function of W and CoFeB thickness. This leads to a value of T = 0.81 ± 0.03 for the ß-W/CoFeB interface. A stark variation of Geff and T with the thickness of the W layer is obtained in accordance with the structural phase transition and resistivity variation of W with its thickness. Effects such as spin memory loss and two-magnon scattering are found to have minor contributions to damping modulation in comparison to the spin pumping effect which is reconfirmed from the unchanged damping constant with the variation of Cu spacer layer thickness inserted between W and CoFeB. The giant interfacial spin transparency and its strong dependence on crystal structures of W will be important for future spin-orbitronic devices based on pure spin current.

9.
Int J Biol Macromol ; 163: 2259-2269, 2020 Nov 15.
Article in English | MEDLINE | ID: mdl-32961193

ABSTRACT

Understanding of interactions of nanomaterials with biomolecules (especially proteins) is of great importance to the area of nanobiotechnology. Graphene and its derivative such as graphene oxide (GO), are two-dimensional (2-D) nanomaterials with remarkable physical and chemical properties and have been broadly explored in biotechnology and biomedical application. Here, we have reported the nature of adsorption of trypsin on the GO surface, considering its biomedical implications. A simple incubation of trypsin on GO surface exhibits varying resistance to autolysis. The structural morphology of trypsin on the GO surface was studied by using atomic force microscopy (AFM), circular dichroism (CD), fluorescence, and total internal reflection fluorescence (TIRF) microscopies. Results suggest that the trypsin follows the Freundlich Isotherm. By the Langmuir model, the maximum adsorption capacity was found to be 100 mg/g. From protein assay results we have concluded that the native trypsin exhibits the highest catalytic efficiency (33.97*104 L mol-1 min-1) in comparison to other Trp-GO constructs. We have further visualized morphological change on GO-trypsin interface throughout the adsorption process by taking samples at definite time intervals, which suggests that the interaction of trypsin with GO is an example of the soft corona. Our findings may be implicated in enzyme engineering as well as enzyme-based bio-sensing applications.


Subject(s)
Graphite/chemistry , Nanostructures/chemistry , Protein Corona/chemistry , Trypsin/chemistry , Adsorption/drug effects , Catalysis , Oxides
11.
Biomolecules ; 10(5)2020 05 04.
Article in English | MEDLINE | ID: mdl-32375303

ABSTRACT

Recent advances in woundcare is targeted towards developing active-dressings, where multiple components are combined to provide a suitable environment for rapid healing. The aim of the present research is to study the preparation of biomimic composite wound dressings by the grafting of hydrogel on silk fibroin fabric. The swelling ability of hydrogel grafted silk fibroin fabric was optimized by changing the initiator concentration. In order to impart antimicrobial properties, these dressing are further coated sono-chemically with zinc oxide nanoparticles. The water vapor transmission rate of the prepared samples was measured. The conformation of silk fibroin proteins after grafting with hydrogel was also confirmed using Fourier Transform Infrared Spectroscopy (FTIR). The morphology of the zinc oxide-coated silk fibroin fabric and hydrogel-coated silk fibroin was studied using Scanning Electron Microscope (SEM). The antimicrobial activity of the zinc oxide-coated samples was studied against E coli. The cytocompatibility of the prepared dressing materials were evaluated using L929 fibroblast cells. MTT assay and phase contrast microscopic studies showed that the adherence, growth, and proliferation of the L929 fibroblast cells that were seeded on zinc oxide nanoparticles on the functionalized hydrogel-coated silk fibroin dressing was significantly higher than that of pure silk fibroin due to the highly porous, bio-mimic structure that allowed ease of passage of nutrients, growth factors, metabolites, and the exchange of gases which is beneficial for successful regeneration of damaged tissues. The expression of TNF-α and IL-2 were not significantly higher than that of control. The proposed composite dressing would be a promising material for wound dressing and regenerative medicine but in order to prove the efficacy of these materials, more in vivo experiments and clinical tests are required to be conducted in future.


Subject(s)
Bandages, Hydrocolloid , Fibroins/chemistry , Metal Nanoparticles/chemistry , Nanocomposites/chemistry , Zinc Oxide/chemistry , Animals , Anti-Infective Agents/chemistry , Anti-Infective Agents/pharmacology , Cell Line , Cytokines/metabolism , Escherichia coli/drug effects , Fibroblasts/drug effects , Fibroblasts/metabolism , Hydrogels/chemistry , Mice , Nanocomposites/adverse effects
12.
ACS Omega ; 5(10): 5041-5047, 2020 Mar 17.
Article in English | MEDLINE | ID: mdl-32201790

ABSTRACT

Graphene oxide-silver nanocomposite (GO-Ag) was fabricated via the sonochemical method, which shows unique physiochemical properties. Graphene oxide (GO) and silver nanoparticles (AgNPs) were synthesized by modified Hummer's and Chemical reduction methods, respectively. The synthesized nanocomposite was characterized using powder X-ray diffraction, Raman spectroscopy, and Fourier-transform infrared spectroscopy. The surface morphology of synthesized nanoparticles was studied using scanning electron microscopy and transmission electron microscopy. The thermoluminescence property of the nanocomposite was analyzed by irradiating the samples in gamma radiation at 1 kGy. Electrochemical reversibility of the GO-Ag nanocomposite was examined by cyclic voltammetry. The photocatalytic application of the nanocomposite was studied using degradation of methylene blue dye. Results reveal that doping of AgNPs on the GO surface not only improves its dye degradation property but also enhances its thermoluminescence property. This knowledge will be helpful in determining the antibacterial property of the GO-Ag nanocomposite in the future.

13.
J Mol Graph Model ; 75: 233-240, 2017 08.
Article in English | MEDLINE | ID: mdl-28600973

ABSTRACT

ß-trefoil is one of the superfolds among proteins. Important classes of proteins like Interleukins (ILs), FibroblastGrowth Factors (FGFs), Kunitz (STI) family of inhibitors etc. belong to this fold. Kunitz (STI) family of inhibitors of proteins possess a highly conserved and structurally important Trytophan 91 (W91) residue, which stitches the top layer of the barrel with the lid. In this article we have investigated the molecular insights of the involvement of this W91 residue in the stability and folding pathway of Kunitz (STI) family. Winged bean Chymotrypsin inhibitor (WCI), a member of Kunitz (STI) family was chosen as a model system for carrying out the work. Molecular dynamics (MD) simulations were run with a set of total six proteins, including wild type WCI (WT) & five mutants namely W91F, W91M, W91A, W91H and W91I. Among all of them the coordinates of four proteins were taken from their crystal structures deposited in the Protein Data Bank (PDB), where as the coordinates for the rest two was generated using in-silico modelling. Our results suggest that truly this W91 residue plays a determining role in stability and folding pathway of Kunitz (STI) family. The mutants are less stable and more susceptible to quicker unfolding at higher temperatures compared to the wild type WCI. These effects are most pronounced for the smallest mutants namely W91H and W91A, indicating more is the cavity created by mutation at W91 position more the proteins becomes unstable.


Subject(s)
Conserved Sequence , Models, Molecular , Plant Proteins/chemistry , Plant Proteins/metabolism , Tryptophan/chemistry , Amino Acid Sequence , Enzyme Stability , Internet , Molecular Dynamics Simulation , Mutant Proteins/chemistry , Mutation/genetics , Principal Component Analysis , Protein Folding , Structure-Activity Relationship
14.
Chem Biol Interact ; 254: 221-30, 2016 Jul 25.
Article in English | MEDLINE | ID: mdl-27270449

ABSTRACT

BACKGROUND: The transition metal oxide nanoparticles are in focus for their anti-cancer potential. In this study we have synthesized and characterized CuO, NiO and Fe2O3 nanoparticles and, investigated their cytotoxic potential in the heterogeneous tumour microenvironment. METHODS: Nanoparticles were synthesized by aqueous precipitation method and characterized with UV-Visible spectrophotometer, Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM) and X-ray diffraction (XRD). Cell viability of lung cancer cells (A549) grown in normoxia (18%O2) and hypoxia (1%O2) was determined for all nanoparticles. The mechanism of cell death was assessed by nuclear morphological analysis, flow cytometry analysis and western blotting. Generation of intracellular ROS in treated cells and its contribution to cell viability was determined. RESULTS: The synthesized metal oxide nanoparticles were successfully characterized with SEM, spectroscopy and X-ray diffraction patterns. Cell viability of lung cancer cells was compromised in both normoxia and hypoxia. ROS generation was shown to contribute to cellular toxicity in CuO, but not NiO and Fe2O3. CONCLUSION: We have shown the therapeutic potential of CuO, NiO and Fe2O3 nanoparticles in non small cell lung cancer cells cultured in hypoxia, a relevant feature of solid tumors along with normoxia. The newly synthesized nanoparticles showed efficacy in both conditions. GENERAL SIGNIFICANCE: Hypoxia drives metabolic alterations and epigenetic modifications in the tumor microenvironment. By using conditions that mimic tumour microenvironment, this study expands the possibility of using metal oxide nanoparticles as a therapeutic agent for lung cancer treatment.


Subject(s)
Apoptosis/drug effects , Metal Nanoparticles , Oxides/pharmacology , Transition Elements/pharmacology , Tumor Microenvironment , A549 Cells , Antineoplastic Agents/chemistry , Antineoplastic Agents/pharmacology , Blotting, Western , Cell Survival/drug effects , Flow Cytometry , Histones/metabolism , Humans , Lung Neoplasms/metabolism , Lung Neoplasms/pathology , Microscopy, Electron, Scanning , Poly (ADP-Ribose) Polymerase-1/metabolism , Reactive Oxygen Species/metabolism , Spectroscopy, Fourier Transform Infrared , Surface Plasmon Resonance , Thermogravimetry , Transition Elements/chemistry , X-Ray Diffraction
15.
Biochim Biophys Acta ; 1854(1): 55-64, 2015 Jan.
Article in English | MEDLINE | ID: mdl-25448016

ABSTRACT

ß-trefoil fold, consisting of a six stranded ß-barrel capped at one end by a lid comprising of another six ß-strands, is one of the most important folds among proteins. Important classes of proteins like Interleukins (ILs), Fibroblast Growth Factors (FGFs), Kunitz (STI) family of inhibitors etc. belong to this fold. Their core is packed by hydrophobic residues contributed by the 6 stranded ß-barrel and three ß-hairpins that make essential contacts with each other and keep the protein in 'topologically minimal frustrated state'. A complete database analysis of the core residues of the ß-trefoil fold proteins presented here identified a conserved tryptophan (W91) residue in the Kunitz (STI) family of inhibitors that projects from the lid and interacts with the bottom layer residues of the barrel. This kind of interactions is unique in Kunitz (STI) family because no other families of ß-trefoil fold have such a shear sized residue at the barrel lid junction; suggesting its possible importance in packing and stability. We took WCI as a representative of this family and prepared four cavity creating mutants W91F-WCI, W91M-WCI, W91I-WCI & W91A-WCI. CD experiments show that the secondary structure of the mutants remains indistinguishable with the wild type. Crystal structures of the mutants W91F-WCI, W91M-WCI & W91A-WCI also show the same feature. However, slight readjustments of the side chains around the site of mutation have been observed so as to minimize the cavity created due to mutation. Comparative stability of these mutants, estimated using heat denaturation CD spectroscopy, indicates that stability of the mutants inversely correlates with the size of the cavity inside the core. Interestingly, although we mutated at the core, mutants show varying susceptibility against tryptic digestion that grossly follow their instability determined by CD. Our findings suggest that the W91 residue plays an important role in determining the stability and packing of the core of WCI.


Subject(s)
Protein Structure, Secondary , Protein Structure, Tertiary , Trypsin Inhibitor, Kunitz Soybean/chemistry , Tryptophan/chemistry , Amino Acids/chemistry , Amino Acids/genetics , Amino Acids/metabolism , Circular Dichroism , Crystallography, X-Ray , Fibroblast Growth Factors/chemistry , Fibroblast Growth Factors/metabolism , Hydrogen Bonding , Hydrophobic and Hydrophilic Interactions , Interleukins/chemistry , Interleukins/metabolism , Models, Molecular , Mutation , Protein Binding , Protein Stability , Temperature , Trypsin Inhibitor, Kunitz Soybean/metabolism , Trypsin Inhibitors/chemistry , Trypsin Inhibitors/metabolism , Tryptophan/genetics , Tryptophan/metabolism
16.
Biochim Biophys Acta ; 1824(7): 882-90, 2012 Jul.
Article in English | MEDLINE | ID: mdl-22709512

ABSTRACT

Canonical serine protease inhibitors interact with cognate enzymes through the P3-P2' region of the inhibitory loop while its scaffold hardly makes any contact. Neighboring scaffolding residues like Arginines or Asparagine shape-up the inhibitory loop and favor the resynthesis of cleaved scissile bond. However, role of remote scaffolding residues, which are not involved in religation, was not properly explored. Crystal structures of two engineered winged bean chymotrypsin inhibitor (WCI) complexed with Bovine trypsin (BPT) namely L65R-WCI:BPT and F64Y/L65R-WCI:BPT show that the inhibitory loop of these engineered inhibitors are recognized and rigidified properly at the enzyme active site like other strong trypsin inhibitors. Chimeric protein ETI(L)-WCI(S), having a loop of Erythrina caffra Trypsin Inhibitor, ETI on the scaffold of WCI, was previously shown to behave like substrate. Non-canonical structure of the inhibitory loop and its flexibility are attributed to the presence of smaller scaffolding residues which cannot act as barrier to the inhibitory loop like in ETI. Double mutant A76R/L115Y-(ETI(L)-WCI(S)), where the barrier is reintroduced on ETI(L)-WCI(S), shows regaining of inhibitory activity. The structure of A76R/L115Y-(ETI(L)-WCI(S)) along with L65R-WCI:BPT and F64Y/L65R-WCI:BPT demonstrate here that the lost canonical conformation of the inhibitory loop is fully restored and loop flexibility is dramatically reduced. Therefore, residues at the inhibitory loop interact with the enzyme playing the primary role in recognition and binding but scaffolding residues having no direct interaction with the enzyme are crucial for rigidification event and the inhibitory potency. B-factor analysis indicates that the amount of inhibitory loop rigidification varies between different inhibitor families.


Subject(s)
Mutant Chimeric Proteins/chemistry , Plant Proteins/chemistry , Trypsin/chemistry , Amino Acid Sequence , Amino Acid Substitution , Animals , Cattle , Crystallography, X-Ray , Escherichia coli/genetics , Models, Molecular , Molecular Sequence Data , Mutant Chimeric Proteins/genetics , Mutation , Plant Proteins/genetics , Protein Engineering , Protein Structure, Secondary , Sequence Alignment
17.
Protein Sci ; 19(3): 593-602, 2010 Mar.
Article in English | MEDLINE | ID: mdl-20073082

ABSTRACT

For canonical serine protease inhibitors (SPIs), scaffolding spacer residue Asn or Arg religates cleaved scissile peptide bond to offer efficient inhibition. However, several designed "mini-proteins," containing the inhibitory loop and the spacer(s) with trimmed scaffold behave like substrates, indicating that scaffolding region beyond the spacer is also important in the inhibitory process. To understand the loop-scaffold compatibility, we prepared three chimeric proteins ECI(L)-WCI(S), ETI(L)-WCI(S), and STI(L)-WCI(S), where the inhibitory loop of ECI, ETI, and STI is placed on the scaffold of their homolog WCI. Results show that although ECI(L)-WCI(S) and STI(L)-WCI(S) behave like good inhibitors, ETI(L)-WCI(S) behaves like a substrate. That means a set of loop residues (SRLRSAFI), offering strong trypsin inhibition in ETI, act as a substrate when they seat on the scaffold of WCI. Crystal structure of ETI(L)-WCI(S) shows that the inhibitory loop is of noncanonical conformation. We identified three novel scaffolding residues Trp88, Arg74, and Tyr113 in ETI that act as barrier to confine the inhibitory loop to canonical conformation. Absence of this barrier in the scaffold of WCI makes the inhibitory loop flexible in ETI(L)-WCI(S) leading to a loss of canonical conformation, explaining its substrate-like behavior. Incorporation of this barrier back in ETI(L)-WCI(S) through mutations increases its inhibitory power, supporting our proposition. Our study provides structural evidence for the contribution of remote scaffolding residues in the inhibitory process of canonical SPIs. Additionally, we rationalize why the loop-scaffold swapping is not permitted even among the members of highly homologous inhibitors, which might be important in the light of inhibitor design.


Subject(s)
Aprotinin/chemistry , Aprotinin/pharmacology , Serine Proteinase Inhibitors/chemistry , Serine Proteinase Inhibitors/pharmacology , Amino Acid Sequence , Aprotinin/genetics , Molecular Sequence Data , Protein Structure, Secondary , Sequence Analysis, Protein , Serine Proteinase Inhibitors/genetics
SELECTION OF CITATIONS
SEARCH DETAIL
...