title: A solid-state NMR study of the interaction of fish antifreeze proteins with phospholipid membranes.
abstract: Fish antifreeze proteins and glycoproteins (AF(G)Ps) prevent ice crystal growth and are able to protect mammalian cells and tissues from hypothermic damage in the sub-zero Polar oceans. This protective mechanism is not fully understood, and further data is required to explain how AF(G)Ps are able to stabilize lipid membranes as they pass through their phase transition temperatures. Solid-state NMR spectroscopy was used as a direct method to study the interaction of the 37-residue alpha-helical type I AFP, TTTT, and the low molecular weight fraction glycoprotein, AFGP8, with dimyristoylphosphatidylcholine membranes above and below the gel-fluid phase transition temperature. In contrast to previous studies in fluid phase bilayers these experiments have provided direct information regarding both the mobility of the phosphate headgroups and perturbation of the acyl chains at a range of temperatures under identical conditions on the same sample. At 5 degrees C changes in the 2H and 31P spectra and a dramatic increase in the 31P T1 relaxation times were consistent with a significant disruption of the membrane by TTTT. Heating to 30 degrees C appeared to expel the peptide from the lipid and re-cooling showed that the interaction of TTTT was not reversible. By contrast, 31P spectra of the membranes with AFGP8 were consistent with interaction with the phosphate headgroups at both 5 and 30 degrees C. Although both peptides interact with the phospholipid bilayer surface, which may stabilize the membrane at lower temperatures, the longer 31P T1 values and the 2H NMR data obtained for TTTT compared with AFGP8 suggest that TTTT causes a greater reduction of phosphate headgroup mobility and has a greater effect on the lipid acyl chains at 5 degrees C.
authors.affiliation: School of Chemistry, The University of New South Wales, Sydney, NSW, 2052, Australia.
authors.forename: James
authors.initials: J
authors.lastname: Garner
authors.forename: Steven R
authors.initials: SR
authors.lastname: Inglis
authors.forename: James
authors.initials: J
authors.lastname: Hook
authors.forename: Frances
authors.initials: F
authors.lastname: Separovic
authors.forename: Margaret M
authors.initials: MM
authors.lastname: Harding
mesh_terms: D000818:Animals; D021301:Antifreeze Proteins / Q000737:chemistry*; D005399:Fishes / Q000378:metabolism*; D008051:Lipid Bilayers / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008560:Membrane Fluidity; D010743:Phospholipids / Q000737:chemistry*; D011485:Protein Binding
chemical_list: D021301:Antifreeze Proteins; D008051:Lipid Bilayers; D010743:Phospholipids