Structural differences of proteins between solution state and solid state probed by attenuated total reflection Fourier transform infrared spectroscopy.
A Fourier transform infrared (FT-IR) spectroscopic method combined with an attenuated total reflection (ATR) sampling technique has been developed to analyze protein secondary structure in both solid and solution states. The method has been applied to analyze the protein structural differences between solution state and solid state. For alpha-helix dominant proteins, beta-sheet structures increase significantly in the solid state, with significant decrease in alpha-helical structures. For beta-sheet dominant proteins, beta-sheet structures increase only moderately in the solid state. When proteins are re-dissolved in solution, their structures are re-natured to their native structures, as suggested by the fact that their structures in solution state are similar to those determined by X-ray crystallography or other spectroscopic methods in solution state. The ATR sampling technique avoids the high pressure and chemicals that are needed for the conventional potassium bromide (KBr) disc method for solid samples in FT-IR spectroscopy. Our approach from this study demonstrated that ATR sampling is more appropriate for analysis of protein structures in the solid state.
Department of Chemistry and Biochemistry, University of Massachusetts Dartmouth, North Dartmouth, Massachusetts 02747, USA.
Amita
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Sachdeva
Shuowei
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Cai
D002256:Carbonic Anhydrases / Q000737:chemistry; D002918:Chymotrypsin / Q000737:chemistry; D002919:Chymotrypsinogen / Q000737:chemistry; D003208:Concanavalin A / Q000737:chemistry; D018360:Crystallography, X-Ray; D005612:Freeze Drying; D009113:Muramidase / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry; D017433:Protein Structure, Secondary; D011506:Proteins / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000737:chemistry; D012996:Solutions / Q000737:chemistry; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods*; D014867:Water / Q000737:chemistry
D009211:Myoglobin; D011506:Proteins; D012996:Solutions; D014867:Water; D003208:Concanavalin A; D012710:Serum Albumin, Bovine; D002919:Chymotrypsinogen; D009113:Muramidase; D002918:Chymotrypsin; D002256:Carbonic Anhydrases