Optimization of the hydrochloric acid concentration used for trifluoroacetate removal from synthetic peptides

Andrushchenko, Valery; Vogel, Hans J.; Prenner, Elmar J.

doi:10.1002/psc.793

Cited by 92 publications

(84 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A common procedure is to use an excess of an acid stronger than TFA, such as HCl, reprotonating TFacetate to form the free acid, which is easily removed by freeze-drying (Andrushchenko et al 2007;Sani et al 2007;Beck et al 2001). Since a very strong acid solution may damage the peptide, we chose an ion-exchange procedure, which ensures efficient removal of TFA under mild acid conditions.…”

Section: Introductionmentioning

confidence: 99%

Efficacy and toxicity of the antimicrobial peptide M33 produced with different counter-ions

et al. 2011

View full text Add to dashboard Cite

The tetra-branched peptide M33 (Pini et al. in FASEB J 24:1015-1022, 2010) is under evaluation in animal models for its activity as antimicrobial agent in lung infections and sepsis. The preclinical development of a new drug requires medium-scale manufacture for tests of efficacy, biodistribution, pharmacokinetics and toxicity. In order to produce the most suitable peptide form for these purposes, we evaluated the behaviour of the peptide M33 obtained with different counter-ions. We compared activity and toxicity in vitro and in vivo of the peptide M33 produced as trifluoroacetate salt (TFacetate) and as acetate salt. The two forms did not differ substantially in terms of efficacy in vitro or in vivo but showed different toxicities for human cells and in animals. M33-TFacetate proved to be 5-30% more toxic than M33-acetate for cells derived from normal bronchi and cells carrying ΔF508 mutation in the CFTR gene, the most frequent variant in cystic fibrosis. M33-TFacetate produced manifest signs of in vivo toxicity immediately after administration, whereas M33-acetate only generated mild signs, which disappeared within a few hours. The peptide M33-acetate proved more suitable for the development of a new drug, and was therefore chosen for further characterization.

show abstract

Section: Introductionmentioning

confidence: 99%

Efficacy and toxicity of the antimicrobial peptide M33 produced with different counter-ions

et al. 2011

View full text Add to dashboard Cite

show abstract

“…This is a peptide derived from the homeodomain of the Drosophila The calcein and acrylamide were obtained from Sigma Aldrich. replacing TFA counter-ions with chloride ions [27]. A with an emission and excitation slit of 2.5 nm.…”

mentioning

confidence: 99%

The role of tryptophans on the cellular uptake and membrane interaction of arginine-rich cell penetrating peptides

Jobin

Blanchet

Henry

et al. 2015

Biochimica et Biophysica Acta (BBA) - Biomembranes

View full text Add to dashboard Cite

Cell-penetrating peptides (CPP) are able to efficiently transport cargos across cell membranes without being cytotoxic to cells, thus present a great potential in drug delivery and diagnosis. While the role of cationic residues in CPPs has been well studied, that of Trp is still not clear. Herein 7 peptide analogs of RW9 (RRWWRRWRR, an efficient CPP) were synthesized in which Trp were systematically replaced by Phe residues. Quantification of cellular uptake reveals that substitution of Trp by Phe strongly reduces the internalization of all peptides despite the fact that they strongly accumulate in the cell membrane. Cellular internalization and biophysical studies show that not only the number of Trp residues but also their positioning in the helix and the size of the hydrophobic face they form are important for their internalization efficacy, the highest uptake occurring for the analog with 3 Trp residues. Using CD and ATR-FTIR spectroscopy we observe that all peptides became structured in contact with lipids, mainly in α-helix. Intrinsic tryptophan fluorescence studies indicate that all peptides partition in the membrane in about the same manner (Kp~10(5)) and that they are located just below the lipid headgroups (~10 Å) with slightly different insertion depths for the different analogs. Plasmon Waveguide Resonance studies reveal a direct correlation between the number of Trp residues and the reversibility of the interaction following membrane washing. Thus a more interfacial location of the CPP renders the interaction with the membrane more adjustable and transitory enhancing its internalization ability.

show abstract

“…Peptide fractions from RP-HPLC were pooled and lyophilized to remove acetonitrile. TFA was then removed from the peptide samples by three rounds of acidification (2 mM HCl) and lyophilization as described by Andrushchenko et al [20] to remove TFA. The peptide was then resuspended in 100 % D2O, the pH adjusted to 7.0 and 10% of dimethyl sulfoxide (DMSO) added to obtain a final volume of 150 μl at 660 μM peptide concentration.…”

Section: Nmr Experiments and Formation Of Amyloid Fibrils Of Kelckav mentioning

confidence: 99%

An Amyloidogenic Sequence at the N-Terminus of the Androgen Receptor Impacts Polyglutamine Aggregation

Oppong

Stier

Gaal

et al. 2017

Preprint

View full text Add to dashboard Cite

Abstract:The human androgen receptor (AR) is a ligand inducible transcription factor harboring an amino terminal domain (AR-NTD) hosting the ligand independent activation function. AR-NTD is intrinsically disordered and display aggregation properties conferred by the presence of a poly-glutamine (polyQ) sequence of 22 residues. The length of the polyQ sequence, as well as the presence of adjacent sequence motifs modulate this aggregation property. AR-NTD contains also a conserved sequence motif KELCKAVSVSM that displays an intrinsic property to form amyloid fibrils under mild oxidative conditions of its conserved cysteine residue. As peptide sequences with intrinsic ability to oligomerize are reported to have an impact on the aggregation of polyQ tract, we determined the effect of the KELCKAVSVSM on the polyQ stretch in the context of the AR NTD, using Atomic Force Microscopy (AFM). Here, we present evidence for a crosstalk between the amyloidogenic properties of the KELCKAVSVSM motif and the polyQ stretch at the AR NTD.

show abstract

Optimization of the hydrochloric acid concentration used for trifluoroacetate removal from synthetic peptides

Cited by 92 publications

References 38 publications

Efficacy and toxicity of the antimicrobial peptide M33 produced with different counter-ions

Efficacy and toxicity of the antimicrobial peptide M33 produced with different counter-ions

The role of tryptophans on the cellular uptake and membrane interaction of arginine-rich cell penetrating peptides

An Amyloidogenic Sequence at the N-Terminus of the Androgen Receptor Impacts Polyglutamine Aggregation

Contact Info

Product

Resources

About