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1.
Commun Biol ; 5(1): 1349, 2022 Dec 08.
Article in English | MEDLINE | ID: mdl-36482075

ABSTRACT

Healthy regeneration of tissue relies on a well-orchestrated release of growth factors. Herein, we show the use of synthetic glycosaminoglycans for controlled binding and release of growth factors to induce a desired cellular response. First, we screened glycosaminoglycans with growth factors of interest to determine kon (association rate constant), koff (dissociation rate constant), and Kd (equilibrium rate constant). As proof-of-concept, we functionalized an elastin-like recombinamer (ELR) hydrogel with a synthetic glycosaminoglycan and immobilized fibroblast growth factor 2 (FGF2), demonstrating that human umbilical vein endothelial cells cultured on top of ELR hydrogel differentiated into tube-like structures. Taking this concept further, we developed a tunable macroporous ELR cryogel material, containing a synthetic glycosaminoglycan and FGF2 that showed increased blood vessel formation and reduced immune response compared to control when implanted in a subcutaneous mouse model. These results demonstrated the possibility for specific release of desired growth factors in/from a modular 3D scaffold in vitro and in vivo.


Subject(s)
Endothelial Cells , Glycosaminoglycans , Humans , Animals , Mice , Delayed-Action Preparations
2.
Mater Today Bio ; 16: 100400, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36060106

ABSTRACT

Understanding the interplay between order and disorder in intrinsically disorder proteins (IDPs), and its impact on the properties and features of materials manufactured from them, is a major challenge in the design of protein-based synthetic polymers intended for advanced functions. In this paper an elastin-like diblock co-recombinamer amphiphile (Phe-ELR) based on a hydrophobic block containing five phenylalanine (Phe) residues proximal to the carboxyl function of a glutamic acid (Glu) residue upon folding, and with Glu as the guest residue in the hydrophilic part, was engineered and its assembly behaviour compared with another amphiphilic ELR used as control. Phe-ELR was tailored in order to clarify the impact of the presence of aromatic residues in the amino acid sequence, which even in early studies by Urry's group already demonstrated a certain out-of-trend behaviour compared with other apolar amino acids, especially non-aromatic ones, on ELR behaviour. The combination of several experimental techniques indicates strong molecular interactions associated with the Phe residue, thus resulting in limited reversible character of the temperature-induced transitions during sequential thermal cycles, a lower than expected transition enthalpy, and clear differences in its supramolecular assembly with respect to the control ELR. A distinctive pre-aggregated state for the Phe-ELR under any condition of pH and temperature is found. Eventually, this state gives rise to Phe-core micelles or a solid jelly-like material, depending on the concentration, pH and presence of salts. In conclusion, it appears that the presence of aromatic residues and their ability to promote strong inter- and intramolecular interactions at any temperature and pH causes a complete modification of the order-disorder interplay present in other, non-aromatic ELRs. These molecular events have a profound impact on the physical properties of the resulting polymer when compared with other ELRs. This work helps to shed light on the limits that govern intrinsic disorder in ELRs beyond its inverse temperature transition.

3.
Acta Biomater ; 141: 14-23, 2022 03 15.
Article in English | MEDLINE | ID: mdl-34971785

ABSTRACT

Elastin-like polymers (ELPs) and their chimeric subfamily the silk elastin-like polymers (SELPs) exhibit a lower critical solvation temperature (LCST) behavior in water which has been extensively studied from theoretical, computational and experimental perspectives. The inclusion of silk domains in the backbone of the ELPs effects the molecular dynamics of the elastin-like domains in response to increased temperature above its transition temperature and confers gelation ability. This response has been studied in terms of initial and long-term changes in structures, however, intermediate transition states have been less investigated. Moreover, little is known about the effects of reversible hydration on the elastin versus silk domains in the physical crosslinks. We used spectroscopic techniques to analyze initial, intermediate and long-term states of the crosslinks in SELPs. A combination of thermoanalytical and rheological measurements demonstrated that the fast reversible rehydration of the elastin motifs adjacent to the relatively small silk domains was capable of breaking the silk physical crosslinks. This feature can be exploited to tailor the dynamics of these types of crosslinks in SELPs. STATEMENT OF SIGNIFICANCE: The combination of silk and elastin in a single molecule results in synergy via their interactions to impact the protein polymer properties. The ability of the silk domains to crosslink affects the thermoresponsive properties of the elastin domains. These interactions have been studied at early and late states of the physical crosslinking, while the intermediate states were the focus of the present study to understand the reversible phase-transitions of the elastin domains over the silk physical crosslinking. The thermoresponsive properties of the elastin domains at the initial, intermediate and late states of silk crosslinking were characterized to demonstrate that reversible hydration of the elastin domains influenced the reversibility of the silk crosslinks.


Subject(s)
Elastin , Silk , Elastin/chemistry , Hydrogels/chemistry , Polymers/chemistry , Silk/chemistry , Temperature
4.
Mater Today Bio ; 11: 100119, 2021 Jun.
Article in English | MEDLINE | ID: mdl-34286238

ABSTRACT

Material platforms based on interaction between organic and inorganic phases offer enormous potential to develop materials that can recreate the structural and functional properties of biological systems. However, the capability of organic-mediated mineralizing strategies to guide mineralization with spatial control remains a major limitation. Here, we report on the integration of a protein-based mineralizing matrix with surface topographies to grow spatially guided mineralized structures. We reveal how well-defined geometrical spaces defined within the organic matrix by the surface topographies can trigger subtle changes in single nanocrystal co-alignment, which are then translated to drastic changes in mineralization at the microscale and macroscale. Furthermore, through systematic modifications of the surface topographies, we demonstrate the possibility of selectively guiding the growth of hierarchically mineralized structures. We foresee that the capacity to direct the anisotropic growth of such structures would have important implications in the design of biomineralizing synthetic materials to repair or regenerate hard tissues.

5.
RSC Adv ; 10(59): 35930-35940, 2020 Sep 28.
Article in English | MEDLINE | ID: mdl-35517089

ABSTRACT

The control of the morphology, as well as the physical and chemical properties, of nanopores is a key issue for many applications. Reducing pore size is important in nanopore-based sensing applications as it helps to increase sensitivity. Changes of other physical properties such as surface net charge can also modify transport selectivity of the pores. We have studied how polyelectrolyte layer-by-layer (LBL) surface modification can be used to change the characteristics of nanoporous membranes. Studies were performed with a custom made three-dimensional multilayer microfluidic device able to fit membrane samples. The device allowed us to efficiently control LBL film deposition over blank low-cost commercially available polycarbonate track-etched (PCTE) membranes. We have demonstrated pore diameter reduction and deposition of the layers inside the pores through confocal and SEM images. Posterior impedance measurement studies served to evaluate experimentally the effect of the LBL deposition on the net inner nanopore surface charge and diameter. Measurements using direct current (DC) and alternative current (AC) voltages have demonstrated contrasted behaviors depending on the number and parity of deposited opposite charge layers. PCTE membranes are originally negatively charged and results evidenced higher impedance increases for paired charge LBL depositions. Impedance decreased when an unpaired positive layer was added. These results showed a different influence on the overall ion motility due to the effect of different surface charges. Results have been fit into a model that suggested a strong dependence of nanopores' impedance module to surface charge on conductive buffers, such as Phosphate Buffer Saline (PBS), even on relatively large nanopores. In AC significant differences between paired and unpaired charged LBL depositions tended to disappear as the total number of layers increased.

6.
Regen Biomater ; 6(1): 21-28, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30740239

ABSTRACT

In the field of tissue engineering the choice of materials is of great importance given the possibility to use biocompatible polymers produced by means of biotechnology. A large number of synthetic and natural materials have been used to this purpose and processed into scaffolds using Electrospinning technique. Among materials that could be used for the fabrication of scaffold and degradable membranes, natural polymers such as collagen, elastin or fibroin offer the possibility to design structures strictly similar to the extracellular matrix (ECM). Biotechnology and genetic engineering made possible the advent of a new class of biopolymers called protein-based polymers. One example is represented by the silk-elastin-proteins that combine the elasticity and resilience of elastin with the high tensile strength of silk-fibroin and display engineered bioactive sequences. In this work, we use electrospinning technique to produce a fibrous scaffold made of the co-recombinamer Silk-ELR. Obtained fibres have been characterized from the morphological point of view. Homogeneity and morphology have been explored using Scanning Electron Microscopy. A thorough study regarding the influence of Voltage, flow rate and distance have been carried out to determine the appropriate parameters to obtain the fibrous mats without defects and with a good distribution of diameters. Cytocompatibility has also been in vitro tested. For the first time we use the co-recombinamer Silk-ELR for the fabrication of a 2.5 angioplasty balloon coating. This structure could be useful as a coated scaffold for the regeneration of intima layer of vessels.

7.
Mater Today Bio ; 2: 100007, 2019 Mar.
Article in English | MEDLINE | ID: mdl-32159144

ABSTRACT

The topic of self-assembled structures based on elastin-like recombinamers (ELRs, i.e., elastin-like polymers recombinantly bio-produced) has released a noticeable amount of references in the last few years. Most of them are intended for biomedical applications. In this review, a complete revision of the bibliography is carried out. Initially, the self-assembly (SA) concept is considered from a general point of view, and then ELRs are described and characterized based on their intrinsic disorder. A classification of the different self-assembled ELR-based structures is proposed based on their morphologies, paying special attention to their tentative modeling. The impact of the mechanism of SA on these biomaterials is analyzed. Finally, the implications of ELR SA in biological systems are considered.

8.
N Biotechnol ; 46: 45-53, 2018 Nov 25.
Article in English | MEDLINE | ID: mdl-30012422

ABSTRACT

With the lack of new chemical antibiotics and increasing pathogen resistance to those available, new alternatives are being explored. Antimicrobial peptides (AMPs) with a broad range of effects, including antibacterial, antifungal, and antiviral actions, have emerged as one of the options. They can be produced by recombinant DNA technology, but the chromatographic methods used for peptide purification are expensive and time consuming. Here, we describe the design, production, purification and assessment of the antibacterial activity of the human peptide hepcidin, using an elastin-like recombinamer as fusion partner. The recombinant protein Hep-A200 was produced in Escherichia coli and purified by a non-chromatographic procedure, exploiting the thermal properties of the A200 elastin-like recombinamer. Recombinant Hep-A200 was found to retain antibacterial activity against Gram-positive and Gram-negative species.


Subject(s)
Anti-Bacterial Agents/metabolism , DNA, Recombinant/metabolism , Elastin/metabolism , Hepcidins/biosynthesis , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Bacillus subtilis/drug effects , Cell Proliferation/drug effects , Cell Survival/drug effects , Cells, Cultured , DNA, Recombinant/chemistry , Elastin/chemistry , Escherichia coli/chemistry , Escherichia coli/drug effects , Escherichia coli/metabolism , Hepcidins/chemistry , Hepcidins/pharmacology , Humans , Microbial Sensitivity Tests , Particle Size , Pseudomonas aeruginosa/drug effects , Staphylococcus aureus/drug effects , Surface Properties
9.
J Biomed Mater Res A ; 106(4): 924-934, 2018 04.
Article in English | MEDLINE | ID: mdl-29105979

ABSTRACT

Novel thermo-sensitive elastin-like recombinamers (ELRs) containing bioactive molecules were created for use as a biomimetic biomaterial for tissue regeneration. For effective use for in vivo applications, it is essential to ensure that they do not induce adverse inflammatory, immune, or allergic responses that inhibit tissue repair. Therefore, we sought to establish a pre-clinical approach to evaluate biocompatibility in experimental mice using ELRs as a prototype biomaterial. First, we measured in vitro proliferation and cytokine production from BALB/c and C57BL/6 mouse splenocytes incubated with ELRs. Second, we used a rapid, high throughput in vivo approach in which inflammatory cells and cytokines were measured following an intraperitoneal implantation. Lastly, a subchronic in vivo approach was used in which ELRs or positive controls were subcutaneously implanted and the implantation sites were assessed for inflammation and gene expression. We found that ELRs induced mild inflammation and minimal fibrosis compared to the intense response to Vitoss. Additionally, implantation increased antigen-specific antibody titers for both groups and gene expression profiling of the implantation sites revealed the upregulation of inflammation, fibrosis, and wound healing-related genes in ELR and positive control-implanted mice compared to sham controls. These data demonstrate that ELRs appear safe for use in tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 924-934, 2018.


Subject(s)
Biocompatible Materials/pharmacology , Elastin/immunology , Elastin/pharmacology , Animals , Antigens/blood , Cell Proliferation/drug effects , Cytokines/biosynthesis , Elastin/isolation & purification , Female , Fibrosis , Gene Expression Regulation/drug effects , Inflammation/pathology , Mice, Inbred BALB C , Mice, Inbred C57BL , Models, Animal , Prosthesis Implantation
10.
Biomed Mater ; 12(4): 045011, 2017 Jul 12.
Article in English | MEDLINE | ID: mdl-28471353

ABSTRACT

Genetically engineered protein polymers functionalized with bioactive domains have potential as multifunctional versatile materials for biomedical use. The present work describes the fabrication and characterisation of antimicrobial fibre mats comprising the antimicrobial elastin-like recombinamer (ELR) CM4-A200. The CM4-A200 protein polymer derives from the genetic fusion of the ABP-CM4 antimicrobial peptide from Bombyx mori with 200 repetitions of the pentamer VPAVG. This is the first report on non-crosslinked fibre mats fabricated with an antimicrobial ELR stable in solution. Thermal gravimetric analysis of CM4-A200 fibre mats shows one single degradation step at temperatures above 300 °C, with fibres displaying a higher thermal degradation activation. The electrospun CM4-A200 fibres display high antimicrobial activity against Gram-positive and Gram-negative bacteria with no detectable cytotoxic effects against normal human skin fibroblasts and keratinocytes, revealing the great potential of these polymers for the fabrication of biomedical materials.


Subject(s)
Anti-Infective Agents/chemistry , Biomedical Engineering/methods , Bombyx/chemistry , Elastin/chemistry , Fibroblasts/chemistry , Polymers/chemistry , Protein Engineering/methods , Animals , Biocompatible Materials , Humans
11.
Biofabrication ; 8(4): 045009, 2016 11 15.
Article in English | MEDLINE | ID: mdl-27845938

ABSTRACT

Coronary angioplasty is the most widely used technique for removing atherosclerotic plaques in blood vessels. The regeneration of the damaged intima layer after this treatment is still one of the major challenges in the field of cardiovascular tissue engineering. Different polymers have been used in scaffold manufacturing in order to improve tissue regeneration. Elastin-mimetic polymers are a new class of molecules that have been synthesized and used to obtain small diameter fibers with specific morphological characteristics. Elastin-like polymers produced by recombinant techniques and called elastin-like recombinamers (ELRs) are particularly promising due to their high degree of functionalization. Generally speaking, ELRs can show more complex molecular designs and a tighter control of their sequence than other chemically synthetized polymers Rodriguez Cabello et al (2009 Polymer 50 5159-69, 2011 Nanomedicine 6 111-22). For the fabrication of small diameter fibers, different ELRs were dissolved in 2,2,2-fluoroethanol (TFE). Dynamic light scattering was used to identify the transition temperature and get a deep characterization of the transition behavior of the recombinamers. In this work, we describe the use of electrospinning technique for the manufacturing of an elastic fibrous scaffold; the obtained fibers were characterized and their cytocompatibility was tested in vitro. A thorough study of the influence of voltage, flow rate and distance was carried out in order to determine the appropriate parameters to obtain fibrous mats without beads and defects. Moreover, using a rotating mandrel, we fabricated a tubular scaffold in which ELRs containing different cell adhesion sequences (mainly REDV and RGD) were collected. The stability of the scaffold was improved by using genipin as a crosslinking agent. Genipin-ELRs crosslinked scaffolds  show a good stability and fiber morphology. Human umbilical vein endothelial cells  were used to assess the in vitro bioactivity of the cell adhesion domains within the backbone of the ELRs.


Subject(s)
Biocompatible Materials/chemistry , Elastin/chemistry , Nanofibers/chemistry , Tissue Scaffolds/chemistry , Amino Acid Sequence , Atherosclerosis/metabolism , Atherosclerosis/pathology , Atherosclerosis/therapy , Biocompatible Materials/pharmacology , Biocompatible Materials/therapeutic use , Cell Adhesion/drug effects , Cell Survival/drug effects , Cross-Linking Reagents/chemistry , Dynamic Light Scattering , Human Umbilical Vein Endothelial Cells , Humans , Iridoids/chemistry , Microscopy, Electron, Scanning , Polymers/chemistry , Tissue Engineering/methods
12.
Nanoscale Res Lett ; 4(10): 1247-1253, 2009 Jul 16.
Article in English | MEDLINE | ID: mdl-20596391

ABSTRACT

Electrostatic self-assembly was used to fabricate new smart multi-layer coatings, using a recombinant elastin-like polymer (ELP) and chitosan as the counterion macromolecule. The ELP was bioproduced, purified and its purity and expected molecular weight were assessed. Aggregate size measurements, obtained by light scattering of dissolved ELP, were performed as a function of temperature and pH to assess the smart properties of the polymer. The build-up of multi-layered films containing ELP and chitosan, using a layer-by-layer methodology, was followed by quartz-crystal microbalance with dissipation monitoring. Atomic force microscopy analysis permitted to demonstrate that the topography of the multi-layered films could respond to temperature. This work opens new possibilities for the use of ELPs in the fabrication of biodegradable smart coatings and films, offering new platforms in biotechnology and in the biomedical area.

13.
J Biomed Mater Res A ; 78(2): 343-51, 2006 Aug.
Article in English | MEDLINE | ID: mdl-16646066

ABSTRACT

Poly(L-valine-L-proline-L-alanine-L-valine-L-glycine) (VPAVG) is a new kind of proteinaceous polymer belonging to the Elastin-like family. These polymers are based on the recurrence of certain short peptide monomers that are considered as "building blocks" in the natural elastin. This smart thermoresponsive polymer has the ability to self-associate at physiological temperature to form aggregates with about 60% in water. This ability can be harnessed to prepare microparticles loaded with an active substance. The aim of this report is to evaluate, from the results of the experiment conducted, the biocompatibility of microparticles prepared from poly(VPAVG). We have studied the cytotoxic effects of microparticles, edema formation after subcutaneous injection (1 and 2.5 mg) in rats (n = 6), and also intraocular tolerance after the intravitreal injection of 2.5 mg of poly(VPAVG) microparticles into pigmented rabbits (n = 12). The polymer did not induce any cytotoxicity or nonspecific depression of cellular respiration on macrophages under the range of polymer concentrations investigated in this study (20, 30, 40, and 60 mg/mL). We observed no inflammatory response to microparticles after subcutaneous injection in the hind-paw of rats, with no significant differences between the control group (PBS) and experimental groups. Anterior and posterior segment signs were evaluated after intraocular injection of poly(VPAVG) microparticles. Only a few eyes (2/11) of the experimental group presented inflammation signs at day 28 postinjection. Nevertheless, 45% (5/11) of the eyes receiving microparticles showed tractional retinal detachment. The results observed in this work suggested certain fibroblastic activity induced by poly(VPAVG) microparticles after their intraocular injection.


Subject(s)
Biocompatible Materials , Elastin , Macrophages, Peritoneal/cytology , Peptides/chemistry , Peptides/pharmacology , Alanine , Amino Acid Sequence , Animals , Cell Survival/drug effects , Conjunctivitis/prevention & control , Edema/prevention & control , Glycine , Macrophages, Peritoneal/drug effects , Mice , Microscopy, Electron, Scanning , Molecular Weight , Peptides/administration & dosage , Proline , Valine , Vitreous Body/drug effects , X-Ray Diffraction
14.
J Control Release ; 102(1): 113-22, 2005 Jan 20.
Article in English | MEDLINE | ID: mdl-15653138

ABSTRACT

Elastin-like polymers (ELPs) are a new kind of protein-based polymers showing interesting properties in the biomaterial field. This work explored the use of self-assembled poly(VPAVG) micro- and nanoparticles as vehicles for the controlled release of the model drug dexamethasone phosphate (DMP). Poly(VPAVG) has shown to form stable particles with a size below 3 mum as a water or PBS polymer solution was warmed above its transition temperature ( approximately 30 degrees C). Due to the peculiar composition of the monomer, the formation and redissolution of the self-assembled microparticles shows an interesting hysteresis behaviour by which the particles are formed at this temperature but do not redissolve until a strong undercooling of approximately 12-15 degrees C is achieved. Therefore, the particles, once formed, are stable either at room or body temperature. These self-assembled particles are able to encapsulate significant amounts of the model drug when self-assembling was carried out in a co-solution polymer-DMP. The release profiles showed a sustained DMP release for about 30 days. Being the potential of this new polymeric carrier high, further research is being conducted to functionalise the poly(VPAVG) base as a way to induce a stronger polymer-drug binding and, accordingly, a more sustained release.


Subject(s)
Drug Carriers/chemistry , Elastin/chemistry , Polymers/chemistry , Delayed-Action Preparations/chemistry , Delayed-Action Preparations/pharmacokinetics , Drug Carriers/pharmacokinetics , Elastin/pharmacokinetics , Microspheres , Particle Size , Pharmaceutical Vehicles , Polymers/pharmacokinetics
15.
Biopolymers ; 62(3): 150-7, 2001.
Article in English | MEDLINE | ID: mdl-11343284

ABSTRACT

Raman spectra of the elastinlike polypentapeptide poly(GVGVP) were measured in H(2)O and D(2)O as solutions and, after increasing the temperature, as suspensions and sediments. In addition, spectra of the polypentapeptide in the solutions of increasing concentration and in the solid state were also investigated by gradually evaporating the water. Significant changes in band frequencies, intensities, and shapes were found for selected Raman bands in the measured spectra, particularly for the C-H stretching, the glycine CH(2) wagging, and some amide vibrations. The C-H stretching vibrations are influenced predominantly by the presence of water, the glycine CH(2) wagging vibrations are associated with conformational transitions. Three possible types of poly(GVGVP)s in the presence of water were indicated: polymer chains in a relatively extended state in the solution, a beta-spiral structure in the suspension, and irregularly bent chains in the sediment.


Subject(s)
Peptides/chemistry , Elastin/chemistry , Models, Molecular , Oligopeptides/chemistry , Protein Structure, Secondary , Spectrum Analysis, Raman , Thermodynamics
18.
Biopolymers ; 54(4): 282-8, 2000 Oct 05.
Article in English | MEDLINE | ID: mdl-10867636

ABSTRACT

The polypentapeptide of elastin, poly(VPGVG), has become an interesting model polypeptide in understanding the mechanism of protein folding and assembly. Due to its simple amino acid composition and the predominance of apolar side chains, this polymer shows strong hydrophobic-hydration phenomena. This paper explores, by calorimetric methods, the nature and structure of the clathrate-like arrangements that take place, surrounding the apolar side chains of the polymer. The performance of these methods, especially differential scanning calorimetry, has a well-gained reputation. In this work, the development of the clathrate-like structures around this model polymer has been followed from water deficiency to water-excess states. Two main conclusions have been obtained from the data obtained. First, there is an upper limit of about 170 water molecules per pentamer as the number of water molecules required to form all the possible clathrate-like structures. Second, these structures exist as an inhomogeneous population with energies spreading in a significantly broad range, which is likely related to differences in geometrical parameters (bond lengths and angles) of the clathrate structure.


Subject(s)
Elastin/chemistry , Peptides/chemistry , Amino Acid Sequence , Calorimetry, Differential Scanning , In Vitro Techniques , Magnetic Resonance Spectroscopy , Thermodynamics , Water/chemistry
19.
J Pineal Res ; 9(2): 103-12, 1990.
Article in English | MEDLINE | ID: mdl-2126038

ABSTRACT

EGTA, a nonspecific ion chelator, showed very good potential for preventing the loss of serotonin N-acetyl transferase activity (arylamine: acetyl-CoA:N-acetyltransferase, EC 2.3.1.5.) (SNAT) from rat and chicken pineal glands during preincubation at 37 degrees C before substrate addition. SNAT activity was higher than that of the controls when EGTA was present in the homogenization buffer due to the prevention of the loss of activity probably not only during the preincubation period but also during incubation with its substrates. This characteristic of EGTA suggests that its use is suitable in the improved SNAT assay described in this paper since higher SNAT activities were found in both rat and chicken pineal glands than the activities recorded with the usual methods.


Subject(s)
Arylamine N-Acetyltransferase/analysis , Pineal Gland/enzymology , Acetyl Coenzyme A/pharmacology , Animals , Chickens , Circadian Rhythm , Egtazic Acid/pharmacology , Enzyme Stability/drug effects , Female , Light , Male , Methods , Rats , Rats, Inbred Strains
20.
J Pineal Res ; 9(4): 243-9, 1990.
Article in English | MEDLINE | ID: mdl-2128943

ABSTRACT

The effects of some general purpose drugs on the deactivation and activity measurement of the chicken pineal gland enzyme serotonin N-acetyl transferase (EC 2.3.1.5.) were studied. The drugs used were EGTA and two neutral surfactants, Nonidet P40 and Triton X-100. Enzyme activity showed significant variations ranging from 2.8 +/- 1.3 nmol/gland/h when Nonidet P40 was added to the homogenate buffer, to 31.8 +/- 1.7 nmol/gland/h when EGTA was present. This striking variation seemed to be caused by the ability of these compounds to modify the rate of NAT deactivation acting either as accelerating agents, as in the case of the detergents or as braking agent, as in the case of EGTA.


Subject(s)
Arylamine N-Acetyltransferase/drug effects , Egtazic Acid/pharmacology , Pineal Gland/enzymology , Surface-Active Agents/pharmacology , Acetyl Coenzyme A/pharmacology , Animals , Arylamine N-Acetyltransferase/metabolism , Chickens , Female , Octoxynol , Pineal Gland/drug effects , Polyethylene Glycols/pharmacology , Time Factors
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