Quantification of the efficiency of cargo delivery by peptidic and pseudo-peptidic Trojan carriers using MALDI-TOF mass spectrometry |
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Authors: | Baptiste Aussedat |
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Affiliation: | Université Pierre et Marie Curie-Paris6, boite 45, 4 place Jussieu, 75252 Paris cedex 05, France CNRS UMR 7613, Synthèse, Structure et Fonction de Molécules Bioactives, FR2769, France |
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Abstract: | We have measured the efficiencies of two novel pseudo-peptidic carriers and various cell-penetrating peptides (Penetratin, (Arg)9 and the third helix of the homeodomain of Knotted-1) to deliver the same cargo inside cells. The cargo that was studied corresponds to the pseudo-substrate of protein kinase C. Cargo delivery was quantified using a recent method based on isotope labeling and MALDI-TOF MS. Results of cargo delivery were compared to the amounts of free CPP internalized inside cells. The third helix of Knotted gave the best results concerning free CPP cellular uptake. It was also found to be the most efficient carrier. This peptide thus emerges as a new CPP with very promising properties. |
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Keywords: | Acm, acetamidomethyl Boc, tert-butoxycarbonyl BSA, bovine serumalbumin CHCA, α-cyano-4-hydroxycinnamic acid CHO, Chinese hamster ovary CPP, cell-penetrating peptide H-CPP, non-deuterated cell-penetrating peptide D-CPP, deuterated cell-penetrating peptide 2Cl-Cbz, 2-chlorobenzyloxycarbonyl DCC, dicyclohexylcarbodiimide DMEM, Dubelcco's modified eagle medium DIEA, diisopropylethylamine DMF, N, N-dimethylformamide DTT, dithiotreitol EDTA, ethylenediamine tetraacetic acid HATU, N-[(dimethylamino)-1H-1,2,3-triazolo[4,5-b]pyridin-1-ylmethylene]-N-methylmethanaminium hexafluorophosphate N-oxide HBTU, N-[(1H-benzotriazol-1-yl)(dimethylamino)methylene]-N-methylmethanaminium hexafluorophosphate N-oxide HOAt, 1-hydroxy-7-aza-benzotriazole HOBt, 1-hydroxybenzotriazole HPLC, high performance liquid chromatography MALDI-TOF MS, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry MBHA-PS, 4-methylbenzhydrylamine polystyrene 4-MeOBzl, p-methoxybenzyl Npys, 3-nitro-2-pyridinesulphenyl PyAOP, 7-Azabenzotriazol-1-yloxytris(pyrrolidino)phosphonium hexafluorophosphate PKC, protein kinase C PKCi, proteins kinase C inhibitor PyBOP, benzotriazol-1-yloxytris(pyrrolidino)phosphonium hexa-fluorophosphate SEM, standard error of the mean TFA, trifluoroacetamide TFFH, tetramethylfluoroformamidinium hexafluorophosphate THF, tetrahydrofuran |
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