NMR spin–echo studies of hydration properties of the molecular chaperone α-crystallin in the bovine lens

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• 作者：Babizhayev ; Mark A. ; Nikolayev ; Gennady N. ; Goryachev ; Sergey N. ; Bours ; Johan
• 关键词：NMR spin&ndash ; echo ; Bovine lens ; Chaperone ; α ; -Crystallin ; Isoelectric focusing ; Protein hydration
• 刊名：Biochimica et Biophysica Acta - Proteins and Proteomics
• 出版年：2002
• 出版时间：July 29, 2002
• 年：2002
• 卷：1598
• 期：1-2
• 页码：46-54
• 全文大小：303 K

文摘

The water-binding properties of bovine lens α-crystallin, collagen from calf skin and bovine serum albumin (BSA), were investigated with various techniques. The water absorptive capacity was obtained in high vacuum desorption experiments volumetrically, and also gravimetrically in controlled atmosphere experiments. NMR spin–echo technique was used to study the hydration of protein samples and to determine the spin–spin relaxation times (T₂) from the protons of water, absorbed on the proteins. Isolated bovine lenses were sectioned into 11–12 morphological layers (from anterior cortex through nucleus to posterior cortex). Crystallin profiles were obtained for each lens layer using thin-layer isoelectric focusing in polyacrylamide gel (IEF). The water content in relation to dry weight of proteins was measured in individual morphological lens layers. During the water vapor uptake P/P₀