Cell-Penetrating Milk-Derived Peptides with a Non-Inflammatory Profile

Area of Science:

• Biochemistry
• Molecular Biology
• Cell Biology

Background:

• Milk-derived peptides possess known anti-inflammatory properties.
• Cell-penetrating peptides (CPPs) offer potential for intracellular delivery.
• Investigating milk-derived CPPs could reveal novel therapeutic applications.

Purpose of the Study:

• To identify and synthesize cell-penetrating peptides from milk protein Alpha-S1-casein.
• To evaluate the cell penetrability and anti-inflammatory effects of these peptides.
• To explore their potential as carriers for intracellular delivery.

Main Methods:

• Computational prediction using machine learning to identify potential CPPs from Alpha-S1-casein.
• Synthesis of selected milk-derived peptides.
• Assessment of cell penetrability via confocal microscopy using fluorescently labeled peptides.
• Evaluation of anti-inflammatory effects by measuring NF-κB activation and secretion of TNFα and IL-1β in HeLa and J774.2 cell lines.

Main Results:

• Several milk-derived peptides were identified and synthesized, predicted to be cell-penetrating.
• All tested peptides demonstrated cell-penetrating capabilities confirmed by confocal microscopy.
• No significant effects were observed on NF-κB transcription factor activation or the secretion of TNFα and IL-1β.
• The peptides were internalized into cells without disrupting key homeostatic inflammatory pathways.

Conclusions:

• Milk-derived peptides from Alpha-S1-casein can effectively penetrate cells.
• These peptides do not modulate major inflammatory pathways like NF-κB activation.
• They hold promise for further research into bioactivities or as natural carriers for enhanced cellular uptake of other molecules.