Probing protein–membrane interactions is vital
for understanding
biological functionality for various applications such as drug development,
targeted drug delivery, and creation of functional biomaterials for
medical and industrial purposes. In this study, we have investigated
interaction of Human Serum Albumin (HSA) with two different lipids,
dipalmitoylphosphatidylglycerol (dDPPG) and dipalmitoylphosphatidylcholine
(dDPPC), using Vibrational Sum Frequency Generation spectroscopy at
different membrane fluidity values. In the liquid-expanded (LE) state
of the lipid, HSA (at pH 3.5) deeply intercalated lipid chains through
a combination of electrostatic and hydrophobic interactions, which
resulted in more ordering of the lipid chains. However, in the liquid-condensed
(LC) state, protein intercalation is decreased due to tighter lipid
packing. Moreover, our findings revealed distinct differences in HSA’s
interaction with dDPPG and dDPPC lipids. The interaction with dDPPC
remained relatively weak compared to dDPPG. These results shed light
on the significance of protein mediated changes in lipid characteristics,
which hold considerable implications for understanding membrane protein
behavior, lipid-mediated cellular processes, and lipid-based biomaterial
design.