Multiple orientation of melittin inside a single lipid bilayer determined by combined vibrational spectroscopic studies.
Despite the availability of several mature structure determination techniques for bulk proteins, determination of structural and orientational information of interfacial proteins, e.g., in cell membranes or on biomaterial surfaces, remains a difficult problem. We combine sum frequency generation (SFG) vibrational spectroscopy with attenuated total reflection-Fourier transform infrared spectroscopy (ATR-FTIR) to investigate the orientation of alpha-helical peptides reconstituted in substrate supported lipid bilayers. Melittin was chosen as a model for alpha-helical peptides, and its orientation when interacting with a supported 1,2-dipalmitoyl-sn-glycero-3-phosphoglycerol (DPPG) bilayer has been examined. Through polarization analysis using amide I signals obtained from both SFG and ATR-FTIR measurements, the orientation distribution of melittin inside a DPPG bilayer was deduced using several trial distribution functions. Melittin was modeled as either an ideal helix or a helix with a bent structure. It was found that a simple distribution function such as a delta-distribution or a Gaussian distribution was not adequate to describe the melittin orientation distribution inside a DPPG bilayer. Instead, two populations of melittin, corresponding to two melittin-bilayer association states, could be used to interpret the experimentally observed result. The method employed in this study demonstrates the feasibility of acquiring a more accurate orientation distribution of peptides/proteins in situ using a combination of vibrational spectroscopic techniques without exogenous labeling.
Chen
Xiaoyun
X
Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA.
Wang
Jie
J
Boughton
Andrew P
AP
Kristalyn
Cornelius B
CB
Chen
Zhan
Z
D008051:Lipid Bilayers / Q000737:chemistry*; D008555:Melitten / Q000737:chemistry*; D008958:Models, Molecular; D010455:Peptides / Q000737:chemistry; D010715:Phosphatidylglycerols / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013499:Surface Properties
D008051:Lipid Bilayers; D010455:Peptides; D010715:Phosphatidylglycerols; D008555:Melitten; C030345:1,2-dipalmitoylphosphatidylglycerol