Metabolic cleavage of cell-penetrating peptides in contact with epithelial models: human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58)

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Metabolic cleavage of cell-penetrating peptides in contact with epithelial models : human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58). / Tréhin, Rachel; Nielsen, Hanne Mørck; Jahnke, Heinz-Georg; Krauss, Ulrike; Beck-Sickinger, Annette G.; Merkle, Hans P.

In: Biochemical Journal, Vol. 382, No. Pt 3, 2004, p. 945-56.

Research output: Contribution to journalJournal articleResearchpeer-review

Harvard

Tréhin, R, Nielsen, HM, Jahnke, H-G, Krauss, U, Beck-Sickinger, AG & Merkle, HP 2004, 'Metabolic cleavage of cell-penetrating peptides in contact with epithelial models: human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58)', Biochemical Journal, vol. 382, no. Pt 3, pp. 945-56. https://doi.org/10.1042/BJ20040238

APA

Tréhin, R., Nielsen, H. M., Jahnke, H-G., Krauss, U., Beck-Sickinger, A. G., & Merkle, H. P. (2004). Metabolic cleavage of cell-penetrating peptides in contact with epithelial models: human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58). Biochemical Journal, 382(Pt 3), 945-56. https://doi.org/10.1042/BJ20040238

Vancouver

Tréhin R, Nielsen HM, Jahnke H-G, Krauss U, Beck-Sickinger AG, Merkle HP. Metabolic cleavage of cell-penetrating peptides in contact with epithelial models: human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58). Biochemical Journal. 2004;382(Pt 3):945-56. https://doi.org/10.1042/BJ20040238

Author

Tréhin, Rachel ; Nielsen, Hanne Mørck ; Jahnke, Heinz-Georg ; Krauss, Ulrike ; Beck-Sickinger, Annette G. ; Merkle, Hans P. / Metabolic cleavage of cell-penetrating peptides in contact with epithelial models : human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58). In: Biochemical Journal. 2004 ; Vol. 382, No. Pt 3. pp. 945-56.

Bibtex

@article{cecee8b0081b449e942032a6ad203b02,
title = "Metabolic cleavage of cell-penetrating peptides in contact with epithelial models: human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58)",
abstract = "We assessed the metabolic degradation kinetics and cleavage patterns of some selected CPP (cell-penetrating peptides) after incubation with confluent epithelial models. Synthesis of N-terminal CF [5(6)-carboxyfluorescein]-labelled CPP, namely hCT (human calcitonin)-derived sequences, Tat(47-57) and penetratin(43-58), was through Fmoc (fluoren-9-ylmethoxycarbonyl) chemistry. Metabolic degradation kinetics of the tested CPP in contact with three cell-cultured epithelial models, MDCK (Madin-Darby canine kidney), Calu-3 and TR146, was evaluated by reversed-phase HPLC. Identification of the resulting metabolites of CF-hCT(9-32) was through reversed-phase HPLC fractionation and peak allocation by MALDI-TOF-MS (matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry) or direct MALDI-TOF-MS of incubates. Levels of proteolytic activity varied highly between the investigated epithelial models and the CPP. The Calu-3 model exhibited the highest proteolytic activity. The patterns of metabolic cleavage of hCT(9-32) were similar in all three models. Initial cleavage of this peptide occurred at the N-terminal domain, possibly by endopeptidase activity yielding both the N- and the C-terminal counterparts. Further metabolic degradation was by aminopeptidase, endopeptidase and/or carboxypeptidase activities. In conclusion, when in contact with epithelial models, the studied CPP were subject to efficient metabolism, a prerequisite of cargo release on the one hand, but with potential for premature cleavage and loss of the cargo as well on the other. The results, particularly on hCT(9-32), may be used as a template to suggest structural modifications towards improved CPP performance.",
keywords = "Amino Acid Sequence, Animals, Biotransformation, Calcitonin, Cell Line, Chromatography, High Pressure Liquid, Dogs, Drug Carriers, Epithelial Cells, Gene Products, tat, Homeodomain Proteins, Humans, Kinetics, Molecular Sequence Data, Molecular Weight, Peptide Fragments, Structure-Activity Relationship, tat Gene Products, Human Immunodeficiency Virus",
author = "Rachel Tr{\'e}hin and Nielsen, {Hanne M{\o}rck} and Heinz-Georg Jahnke and Ulrike Krauss and Beck-Sickinger, {Annette G.} and Merkle, {Hans P}",
year = "2004",
doi = "10.1042/BJ20040238",
language = "English",
volume = "382",
pages = "945--56",
journal = "Biochemical Journal",
issn = "0264-6021",
publisher = "Portland Press Ltd.",
number = "Pt 3",

}

RIS

TY - JOUR

T1 - Metabolic cleavage of cell-penetrating peptides in contact with epithelial models

T2 - human calcitonin (hCT)-derived peptides, Tat(47-57) and penetratin(43-58)

AU - Tréhin, Rachel

AU - Nielsen, Hanne Mørck

AU - Jahnke, Heinz-Georg

AU - Krauss, Ulrike

AU - Beck-Sickinger, Annette G.

AU - Merkle, Hans P

PY - 2004

Y1 - 2004

N2 - We assessed the metabolic degradation kinetics and cleavage patterns of some selected CPP (cell-penetrating peptides) after incubation with confluent epithelial models. Synthesis of N-terminal CF [5(6)-carboxyfluorescein]-labelled CPP, namely hCT (human calcitonin)-derived sequences, Tat(47-57) and penetratin(43-58), was through Fmoc (fluoren-9-ylmethoxycarbonyl) chemistry. Metabolic degradation kinetics of the tested CPP in contact with three cell-cultured epithelial models, MDCK (Madin-Darby canine kidney), Calu-3 and TR146, was evaluated by reversed-phase HPLC. Identification of the resulting metabolites of CF-hCT(9-32) was through reversed-phase HPLC fractionation and peak allocation by MALDI-TOF-MS (matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry) or direct MALDI-TOF-MS of incubates. Levels of proteolytic activity varied highly between the investigated epithelial models and the CPP. The Calu-3 model exhibited the highest proteolytic activity. The patterns of metabolic cleavage of hCT(9-32) were similar in all three models. Initial cleavage of this peptide occurred at the N-terminal domain, possibly by endopeptidase activity yielding both the N- and the C-terminal counterparts. Further metabolic degradation was by aminopeptidase, endopeptidase and/or carboxypeptidase activities. In conclusion, when in contact with epithelial models, the studied CPP were subject to efficient metabolism, a prerequisite of cargo release on the one hand, but with potential for premature cleavage and loss of the cargo as well on the other. The results, particularly on hCT(9-32), may be used as a template to suggest structural modifications towards improved CPP performance.

AB - We assessed the metabolic degradation kinetics and cleavage patterns of some selected CPP (cell-penetrating peptides) after incubation with confluent epithelial models. Synthesis of N-terminal CF [5(6)-carboxyfluorescein]-labelled CPP, namely hCT (human calcitonin)-derived sequences, Tat(47-57) and penetratin(43-58), was through Fmoc (fluoren-9-ylmethoxycarbonyl) chemistry. Metabolic degradation kinetics of the tested CPP in contact with three cell-cultured epithelial models, MDCK (Madin-Darby canine kidney), Calu-3 and TR146, was evaluated by reversed-phase HPLC. Identification of the resulting metabolites of CF-hCT(9-32) was through reversed-phase HPLC fractionation and peak allocation by MALDI-TOF-MS (matrix-assisted laser-desorption ionization-time-of-flight mass spectrometry) or direct MALDI-TOF-MS of incubates. Levels of proteolytic activity varied highly between the investigated epithelial models and the CPP. The Calu-3 model exhibited the highest proteolytic activity. The patterns of metabolic cleavage of hCT(9-32) were similar in all three models. Initial cleavage of this peptide occurred at the N-terminal domain, possibly by endopeptidase activity yielding both the N- and the C-terminal counterparts. Further metabolic degradation was by aminopeptidase, endopeptidase and/or carboxypeptidase activities. In conclusion, when in contact with epithelial models, the studied CPP were subject to efficient metabolism, a prerequisite of cargo release on the one hand, but with potential for premature cleavage and loss of the cargo as well on the other. The results, particularly on hCT(9-32), may be used as a template to suggest structural modifications towards improved CPP performance.

KW - Amino Acid Sequence

KW - Animals

KW - Biotransformation

KW - Calcitonin

KW - Cell Line

KW - Chromatography, High Pressure Liquid

KW - Dogs

KW - Drug Carriers

KW - Epithelial Cells

KW - Gene Products, tat

KW - Homeodomain Proteins

KW - Humans

KW - Kinetics

KW - Molecular Sequence Data

KW - Molecular Weight

KW - Peptide Fragments

KW - Structure-Activity Relationship

KW - tat Gene Products, Human Immunodeficiency Virus

U2 - 10.1042/BJ20040238

DO - 10.1042/BJ20040238

M3 - Journal article

VL - 382

SP - 945

EP - 956

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - Pt 3

ER -

ID: 43890710