Mixed trehalose/sucrose glasses used for protein incorporation as studied by infrared and optical spectroscopy.
Evaporation of water from a 1/1 mixture of trehalose and sucrose gives rise to optically clear glasses that are transparent in the UV and visible ranges and do not crystallize when they are prepared at ambient temperatures. Two proteins, liver alcohol dehydrogenase and parvalbumin, and the tryptophan derivative N-acetyl-tryptophanamide were incorporated into the glasses. Infrared spectroscopy of the amide I band reveals that the proteins retain secondary structure in the glass over a temperature range of 20-300K. The amide II band of the protein and the HOH bending band of residual water in the glass shift with temperature changes, consistent with increased H-bonding strength as temperature is lowered. Phosphorescence of tryptophan can be seen from the proteins at room temperature, which shows the immobilization of the protein by the glass and the curbing of oxygen diffusion. It is suggested that using mixed sugars to form glasses is a way to immobilize proteins over a wide temperature range without distortions from solvent crystals.
Department of Biochemistry and Biophysics, Johnson Research Foundation, School of Medicine, University of Pennsylvania, Philadelphia, PA 19104, USA.
Wayne W
WW
Wright
Juan
J
Carlos Baez
Jane M
JM
Vanderkooi
D000426:Alcohol Dehydrogenase / Q000737:chemistry; D000818:Animals; D003460:Crystallization; D005399:Fishes; D005612:Freeze Drying; D005898:Glass / Q000737:chemistry*; D006736:Horses; D006860:Hydrogen Bonding; D008099:Liver / Q000201:enzymology; D010100:Oxygen / Q000378:metabolism; D010320:Parvalbumins / Q000737:chemistry; D010766:Phosphorylation; D017433:Protein Structure, Secondary; D013053:Spectrophotometry; D013055:Spectrophotometry, Infrared; D013395:Sucrose / Q000737:chemistry*; D013816:Thermodynamics; D014199:Trehalose / Q000737:chemistry*; D014364:Tryptophan / Q000031:analogs & derivatives* / Q000737:chemistry; D014867:Water / Q000737:chemistry*
D010320:Parvalbumins; D014867:Water; C016952:N-acetyltryptophanamide; D013395:Sucrose; D014364:Tryptophan; D014199:Trehalose; D000426:Alcohol Dehydrogenase; D010100:Oxygen