Molecular insights into α-Lactalbumin-Chlorogenic acid interaction: Peptide design and anti-inflammatory mechanisms.

In this study, we systematically explored the molecular interaction mechanism between α-lactalbumin (ALA) and chlorogenic acid (CA), and designed the functional peptide TQVYW based on key interfacial residues (THR33, GLN54, VAL99, TYR103, and TRP104) identified from ALA through molecular docking with CA. Furthermore, we elucidated the synergistic anti-inflammatory mechanism of the peptide TQVYW in combination with CA. The study showed that ALA binds CA via hydrogen bonds and hydrophobic interactions, forming a stable molten globule. Further studies showed that TQVYW-CA suppressed reactive oxygen species (ROS) and nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) activation, while reducing both apoptosis and inflammasome formation. It decreased nitric oxide (NO) levels from 40.7 to 14.8 μmol/L (p < 0.001), and downregulated tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and interleukin-6 (IL-6) expression, reducing them by 7.08, 13.65, and 14.08fold, respectively (p < 0.001). TQVYW inhibited lipopolysaccharide (LPS)-induced apoptosis and inflammation in RAW264.7 macrophages, synergistically enhancing anti-inflammatory effects of CA. This study reveals the molecular mechanism of natural peptide amplification of CA bioactivity and proposes an innovative strategy for the development of peptide-CA anti-inflammatory functional foods.