Membrane and protein properties of freeze-dried mouse platelets.
Membrane properties and the overall protein secondary structure of freeze-dried trehalose-loaded mouse platelets were studied using steady state fluorescence anisotropy and Fourier transform infrared spectroscopy (FTIR). FTIR results showed that fresh control mouse platelets have a main phase transition at approximately 14 degrees C, whereas, freeze-dried platelets exhibited a main phase transition approximately 12 degrees C. However, the cooperativity of the transition of the rehydrated platelets was greatly enhanced compared to that of control platelets. Anisotropy experiments performed with 1,6 diphenyl-1,3,5 hexatriene (DPH) complemented FTIR results and showed that the lipid order in the core of the membrane was affected by freeze-drying procedures. Similar experiments with trimethyl ammonium 1,6 diphenyl-1,3,5 hexatriene (TMA-DPH), a membrane surface probe, indicated that membrane properties at the membrane/water interface were less affected by freeze-drying procedures than the core of the membrane. Lyophilization did not result in massive protein denaturation, but the overall protein secondary structure was altered, based on in situ assessment of the amide-I and amide-II band profiles. Lyophilization-induced changes to endogenous platelet proteins were further investigated by studying the protein's heat stability. In fresh control platelets, proteins denatured at 42 degrees C, whereas proteins in the rehydrated platelets denatured at 48 degrees C.
Wolkers
Willem F
WF
wfwolkers@ucdavis.edu
Looper
Sheri A
SA
McKiernan
Ariane E
AE
Tsvetkova
Nelly M
NM
Tablin
Fern
F
Crowe
John H
JH
D000818:Animals; D001792:Blood Platelets / Q000737:chemistry* / Q000502:physiology*; D002462:Cell Membrane / Q000737:chemistry* / Q000502:physiology; D004161:Diphenylhexatriene / Q000031:analogs & derivatives* / Q000737:chemistry; D005260:Female; D005454:Fluorescence Polarization / Q000379:methods; D005456:Fluorescent Dyes / Q000737:chemistry; D005612:Freeze Drying / Q000379:methods*; D008297:Male; D008563:Membrane Lipids / Q000737:chemistry; D051379:Mice; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared; D013696:Temperature
D005456:Fluorescent Dyes; D008563:Membrane Lipids; D011506:Proteins; D004161:Diphenylhexatriene; C033628:1-(4-(trimethylamino)phenyl)-6-phenylhexa-1,3,5-triene