Comparison of the influence of humidity and D-mannitol on the organization of tetraethylene glycol-terminated self-assembled monolayers and immobilized antimicrobial peptides.
We report the use of polarization-modulation infrared reflection-absorption spectroscopy (PM-IRRAS) to characterize the effects of relative humidity (RH) and d-mannitol on the conformations of tetraethylene glycol (EG4)-terminated self-assembled monolayers (SAMs) and immobilized antimicrobial peptides (Cecropin P1 and a hybrid of Cecropin A (1-8) and Melittin (1-18)). These results are used to assess the extent to which d-mannitol can substitute for water in promoting conformational states of the SAMs and oligopeptides similar to those induced by hydration. Our measurements reveal a red shift of the COC asymmetric stretching vibration of the EG4-terminated SAMs with increasing humidity, consistent with a transition from a mixed all-trans/helical (7/2 helix) conformation at 0% RH to a predominantly helical conformation at 90% RH. Significantly, under dry conditions, a thin (2 nm in thickness) overlayer of d-mannitol generated the COC spectroscopic signature of the EG4-terminated SAM measured at high humidity. Comparisons of the effects of humidity and d-mannitol on the secondary structure of the two oligopeptides also revealed both to cause the amide I peak positions, which were measured in dry air (and without d-mannitol) to correspond to α-helical conformations, to undergo red-shifts. The magnitudes of the red-shifts, however, were more pronounced for dry d-mannitol than for high RH, with Cecropin P1 and the hybrid peptide exhibiting amide I peak positions under d-mannitol consistent with bulk aqueous solution secondary structures (random and β-sheet, respectively). These results are discussed in the context of prior reports of the tendency of d-mannitol to form glassy states in the absence of water. Overall, the results presented in this paper support the hypothesis that d-mannitol can substitute, in at least some ways, for the influence of water on the conformational states of biologically relevant molecules at interfaces. The results provide guidance for the design of interfaces for water-free biologics.
Department of Chemical and Biological Engineering, University of Wisconsin-Madison , Madison, Wisconsin 53706, United States.
Mohit
M
Goel
E Neil G
EN
Marsh
Zhan
Z
Chen
Nicholas L
NL
Abbott
D000890:Anti-Infective Agents / Q000737:chemistry*; D023181:Antimicrobial Cationic Peptides / Q000737:chemistry; D006813:Humidity; D008353:Mannitol / Q000737:chemistry*; D008555:Melitten / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D017433:Protein Structure, Secondary
D000890:Anti-Infective Agents; D023181:Antimicrobial Cationic Peptides; D010455:Peptides; D008555:Melitten; D008353:Mannitol; C031161:cecropin A; C070016:cecropin P1-LI