Surfaces
immobilized with biological molecules such as peptides
and proteins are widely used in many important applications including
biosensors, medical devices, and food packaging. It was found that
the structures of surface-immobilized peptides control their surface
properties. In this study, we investigated interfacial behaviors of
antimicrobial peptide cecropin P1 (CP1) immobilized onto a maleimide-terminated
self-assembled monolayer (Mal SAM) and a mixed SAM (Mal-OH SAM, a
mixture of maleimide-terminated SAM and hydroxyl-terminated SAM) surface
via C-terminus modified cysteine (CP1c). The surface coverage, secondary
structure, orientation, and antimicrobial activity of immobilized
CP1c were investigated using surface plasmon resonance (SPR), circular
dichroism (CD), sum frequency generation (SFG) vibrational spectroscopy,
dynamic contact antimicrobial assay, and coarse grained molecular
dynamics (MD) simulations. It was found that the surface coverages
of CP1c on the Mal SAM and the mixed Mal-OH SAM are similar. CP1c
on Mal SAM possessed a dominant helical structure with a single orientation
of ∼32° versus surface normal. CP1c on Mal-OH mixed SAM
surface also possessed a dominant helical structure but with multiple
orientations. MD simulation results can be correlated to the experimental
data: the simulation results indicate a narrow distribution of orientations
of CP1c immobilized on Mal SAM, but multiple orientations are sampled
on the more hydrophilic Mal-OH SAM. Even though the surface orientations
of CP1c immobilized on the two SAM surfaces are different, activity
against both Gram-negative and Gram-positive bacteria (Escherichia coli and Staphylococcus
aureus) exhibited similar results for CP1c immobilized
on both SAM surfaces. We believe that this is because the antimicrobial
activity of the surface-immobilized peptides is mainly affected by
the electrostatic interactions between the strong basic N-terminal
residues and the negatively charged bacteria cell wall/cell membrane.