Tuning Thermoresponsive Properties of Cationic Elastin-like Polypeptides by Varying Counterions and Side-Chains

Petitdemange, Rosine; Garanger, Élisabeth; Bataille, Laure; Bathany, Katell; Garbay, Bertrand; Deming, Timothy J.; Lecommandoux, Sébastien

doi:10.1021/acs.bioconjchem.7b00082

Cited by 43 publications

(102 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Bioproduction, isolation and purification of the recombinant ELP with the primary sequence MW[VPGVGVPGMG(VPGVG) 2 ] 10 was achieved following previously reported procedures [55].…”

Section: Methodsmentioning

confidence: 99%

“…Introduction of an alkyne handle at the side chain thioether group of each methionine residue was achieved as previously described [55]. Briefly, 50 mg of ELP was dissolved in 2.5 mL of an acetic acid (AcOH)/HFIP mixture (9:1, v/v) (20 mg/mL).…”

Section: Synthetic Procedures 211 Synthesis Of Elp(-cch)mentioning

confidence: 99%

“…pH-responsive and positively-charged ELPs featuring either primary amine (-NH 2 ) or secondary amine (-NHCH 3 ) units were synthetized in two steps from the recombinant ELP with the sequence MW[VPGVGVPGMG(VPGVG) 2 ] 10 . (Scheme 1) This 17 kDa ELP featuring valine or methionine at the guest residue position (Scheme 1a) can be obtained in good yield by recombinant production in Escherichia coli bacteria and subsequent purification from the cytoplasmic soluble fraction by Inverse Transition Cycling [45,54,55]. The ELP was chemoselectively modified onto the methionine residues' side chain (11 total in the sequence) to introduce either primary or secondary amine groups (Scheme 1b).…”

Section: Synthesis Of Ph-responsive Cationic Elpsmentioning

confidence: 99%

“…The ELP was chemoselectively modified onto the methionine residues' side chain (11 total in the sequence) to introduce either primary or secondary amine groups (Scheme 1b). In the first step, methionine residues were alkylated using glycidyl propargyl ether to position alkyne handles [55] for subsequent coupling in the second step with azido-functionalized amines (i.e. 2aminoethylazide and (2-azidoethyl)methylamine) using copper-catalyzed azide-alkyne cycloaddition (CuAAC) [69,70] (Figures S1 and S2).…”

Section: Synthesis Of Ph-responsive Cationic Elpsmentioning

confidence: 99%

“…ELP scaffolds are first produced recombinantly in Escherichia coli (E. coli) bacterial hosts and subsequently chemoselectively modified to introduce various functionalities. In particular, ELPs containing periodically spaced methionine residues have been engineered and chemoselectively modified at the thioether side chain of each methionine residues [26,46,55]. We believed this approach to be particularly relevant for the generation of positively charged ELPs that can be difficult to produce and purify by Inverse Transition Cycling [56,57] in high yields due to high transition temperatures.…”

mentioning

confidence: 99%

See 4 more Smart Citations

Nucleic acids complexation with cationic elastin-like polypeptides: Stoichiometry and stability of nano-assemblies

Bravo-Anaya

Garbay

Nando-Rodríguez

et al. 2019

Journal of Colloid and Interface Science

View full text Add to dashboard Cite

Positively charged elastin-like polypeptides (ELPs) were synthesized for the compaction of genetic material. A recombinant ELP (VPGXG) 40 with X=V,M (3:1) was post-modified in two steps to introduce chemoselectively either primary or secondary amine pendant groups at each methionine residue. Positively charged ELPs were characterized by SDS-PAGE, size exclusion chromatography, 1 H NMR, potentiometric titrations and dynamic light scattering to assess their purity and determine their degree of functionalization, molecular weight, isoelectric point and thermo-responsive behaviour. Electrostatic complexation between the different ELP derivatives and nucleic acids was studied to determine the stoichiometry of ELP S /nucleic acids complex formation, and to find optimal conditions leading to stable nanoparticles with controlled size and surface potential. The stability of these complexes was investigated in the presence of salts at physiological concentrations and in the presence of surfactant. This study revealed that two regimes of stable nanoparticles in terms of size and charge can be obtained from the electrostatic complexation between the primary amine containing ELP derivative, ELP(-NH 2 ), and plasmid DNA. Resulting complexes were found to be stable to dissociation for charge ratios up to 2.5 under physiological salt concentrations (154 mM NaCl), showing that plasmid DNA was completely condensed by the polycationic ELP and protected against electrolyte-mediated dissociation.

show abstract

“…Bioproduction, isolation and purification of the recombinant ELP with the primary sequence MW[VPGVGVPGMG(VPGVG) 2 ] 10 was achieved following previously reported procedures [55].…”

Section: Methodsmentioning

confidence: 99%

Section: Synthetic Procedures 211 Synthesis Of Elp(-cch)mentioning

confidence: 99%

Section: Synthesis Of Ph-responsive Cationic Elpsmentioning

confidence: 99%

Section: Synthesis Of Ph-responsive Cationic Elpsmentioning

confidence: 99%

mentioning

confidence: 99%

See 3 more Smart Citations

Nucleic acids complexation with cationic elastin-like polypeptides: Stoichiometry and stability of nano-assemblies

Bravo-Anaya

Garbay

Nando-Rodríguez

et al. 2019

Journal of Colloid and Interface Science

View full text Add to dashboard Cite

show abstract

Elastin‐Like Recombinamers: Deconstructing and Recapitulating the Functionality of Extracellular Matrix Proteins Using Recombinant Protein Polymers

Acosta

Quintanilla‐Sierra

Mbundi

et al. 2020

Adv Funct Materials

View full text Add to dashboard Cite

In the development of tissue engineering strategies to replace, remodel, regenerate, or support damaged tissue, the development of bioinspired biomaterials that recapitulate the physicochemical characteristics of the extracellular matrix has received increased attention. Given the compositional heterogeneity and tissue-to-tissue variation of the extracellular matrix, the design, choice of polymer, crosslinking, and nature of the resulting biomaterials are normally depended on intended application. Generally, these biomaterials are usually made of degradable or nondegradable biomaterials that can be used as cell or drug carriers. In recent years, efforts to endow reciprocal biomaterial-cell interaction properties in scaffolds have inspired controlled synthesis, derivatization, and functionalization of the polymers used. In this regard, elastin-like recombinant proteins have generated interest and continue to be developed further owing to their modular design at a molecular level. In this review, the authors provide a summary of key extracellular matrix features relevant to biomaterials design and discuss current approaches in the development of extracellular matrix-inspired elastin-like recombinant protein based biomaterials.

show abstract

Control of Enzyme Reactivity in Response to Osmotic Pressure Modulation Mimicking Dynamic Assembly of Intracellular Organelles

et al. 2023

Self Cite

View full text Add to dashboard Cite

In response to variations in osmotic stress, in particular to hypertonicity associated with biological dysregulations, cells have developed complex mechanisms to release their excess water, thus avoiding their bursting and death. When water is expelled, cells shrink and concentrate their internal bio(macro)molecular content, inducing the formation of membraneless organelles following a liquid–liquid phase separation (LLPS) mechanism. To mimic this intrinsic property of cells, functional thermo‐responsive elastin‐like polypeptide (ELP) biomacromolecular conjugates are herein encapsulated into self‐assembled lipid vesicles using a microfluidic system, together with polyethylene glycol (PEG) to mimic cells’ interior crowded microenvironment. By inducing a hypertonic shock onto the vesicles, expelled water induces a local increase in concentration and a concomitant decrease in the cloud point temperature (Tcp) of ELP bioconjugates that phase separate and form coacervates mimicking cellular stress‐induced membraneless organelle assemblies. Horseradish peroxidase (HRP), as a model enzyme, is bioconjugated to ELPs and is locally confined in coacervates as a response to osmotic stress. This consequently increases local HRP and substrate concentrations and accelerates the kinetics of the enzymatic reaction. These results illustrate a unique way to fine‐tune enzymatic reactions dynamically as a response to a physiological change in isothermal conditions.

show abstract

Tuning Thermoresponsive Properties of Cationic Elastin-like Polypeptides by Varying Counterions and Side-Chains

Cited by 43 publications

References 34 publications

Nucleic acids complexation with cationic elastin-like polypeptides: Stoichiometry and stability of nano-assemblies

Nucleic acids complexation with cationic elastin-like polypeptides: Stoichiometry and stability of nano-assemblies

Elastin‐Like Recombinamers: Deconstructing and Recapitulating the Functionality of Extracellular Matrix Proteins Using Recombinant Protein Polymers

Control of Enzyme Reactivity in Response to Osmotic Pressure Modulation Mimicking Dynamic Assembly of Intracellular Organelles

Contact Info

Product

Resources

About