The behavior of hydration water of protein with the protectant in the view of 1HNMR.
The alterations of the amount of hydration water and the extent of the interaction with hydration water and protein during freezing and after freeze-drying were investigated with or without cryoprotectant using 1HNMR and other methods. Ovalbumin was used as a representative protein that was stable against freezing and drying. Myosin was also used as a typical unstable protein. It was found that myosin had no stable hydration layer compared with ovalbumin. It was considered that this difference might be the major cause of the difference in the stability of both proteins against freezing and drying. With cryoprotectant, the amount of hydration water was decreased and the extent of the interaction of hydration water with protein was increased. The results suggested the effective protectant substituted for part of the hydration water and strengthened the interaction during freezing and drying. It is also possible that the formation of a "quasi-hydration layer" might protect protein against denaturation due to freezing and drying.
Hanafusa
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Inst. of Low Temp. Sci., Hokkaido, Univ., Sapporo, Japan.
D003451:Cryoprotective Agents; D004355:Drug Stability; D005612:Freeze Drying / Q000379:methods*; D009682:Magnetic Resonance Spectroscopy; D009218:Myosins / Q000302:isolation & purification; D010047:Ovalbumin / Q000302:isolation & purification; D011489:Protein Denaturation; D011506:Proteins / Q000302:isolation & purification*; D013696:Temperature; D014867:Water / Q000032:analysis
D003451:Cryoprotective Agents; D011506:Proteins; D014867:Water; D010047:Ovalbumin; D009218:Myosins