Hydration state change of proteins upon unfolding in sugar solutions.


Abstract

Change in hydration number of proteins upon unfolding, Deltan, was obtained from the analysis of thermal unfolding behavior of proteins in various sugar solutions with water activity, a(W), varied. By applying the reciprocal form of Wyman-Tanford equation, Deltan was determined to be 133.9, 124.1, and 139.2 per protein molecule for ribonuclease A at pH=5.5, 4.2, and 2.8, respectively, 201.4 for lysozyme at pH=5.5, and 100.1 for alpha-chymotripnogen A at pH=2.0. Among the sugars tested, reducing sugars gave the lower apparent Deltan as compared with nonreducing sugars probably because of the direct interaction of reducing terminal with amino group of proteins at a high temperature. From the knowledge of Deltan, a new thermodynamic model for protein stability was proposed with explicit consideration for hydration state change of protein upon unfolding. From this model, the contribution of a(W) was proven to be always positive for stabilization of proteins and its effect is not negligible depending on Deltan and a(W). Study holds ProTherm entries: 22921, 22922, 22923, 22924, 22925, 22926, 22927, 22928, 22929, 22930, 22931, 22932, 22933, 22934, 22935, 22936, 22937, 22938, 22939, 22940, 22941, 22942, 22943, 22944, 22945, 22946, 22947, 22948, 22949, 22950, 22951, 22952, 22953, 22954, 22955, 22956, 22957, 22958, 22959, 22960, 22961, 22962, 22963, 22964, 22965, 22966, 22967, 22968, 22969, 22970, 22971, 22972, 22973, 22974, 22975, 22976, 22977, 22978, 22979, 22980, 22981, 22982, 22983, 22984, 22985, 22986, 22987, 22988, 22989, 22990, 22991, 22992, 22993, 22994, 22995, 22996, 22997, 22998, 22999, 23000, 23001, 23002, 23003, 23004, 23005, 23006, 23007, 23008, 23009, 23010, 23011, 23012, 23013, 23014, 23015, 23016, 23017, 23018, 23019, 23020, 23021, 23022, 23023, 23024, 23025, 23026, 23027, 23028, 23029, 23030, 23031, 23032, 23033, 23034, 23035, 23036, 23037, 23038, 23039 Extra Details: Data were obtained in 0 M sucrose Thermal unfolding of protein; Hydration number; Water activity; Free energy for protein stability; Ribonuclease A

Submission Details

ID: sKfHK8YF3

Submitter: Connie Wang

Submission Date: April 24, 2018, 8:54 p.m.

Version: 1

Publication Details
Miyawaki O,Biochim. Biophys. Acta (2007) Hydration state change of proteins upon unfolding in sugar solutions. PMID:17581805
Additional Information

Number of data points 238
Proteins Ribonuclease pancreatic ; Lysozyme C ; Ribonuclease pancreatic ; Chymotrypsinogen A ; Lysozyme C ; Calnexin
Unique complexes 3
Assays/Quantities/Protocols Experimental Assay: Tm details:Additives , pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: dHvH details:Additives , pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: Tm details:Additives sucrose (1.9963 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: dHvH details:Additives sucrose (1.9963 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: Tm details:Additives sucrose (1.5563 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: dHvH details:Additives sucrose (1.5563 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: Tm details:Additives sucrose (1.0801 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: dHvH details:Additives sucrose (1.0801 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: Tm details:Additives sucrose (0.5632 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: dHvH details:Additives sucrose (0.5632 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: Tm details:Additives sucrose (0 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: dHvH details:Additives sucrose (0 M),, pH:2.0, buffers:HCl: 0.01 M ; Experimental Assay: Tm details:Additives , pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: dHvH details:Additives , pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: Tm details:Additives sucrose (1.9963 M),, pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: dHvH details:Additives sucrose (1.9963 M),, pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: Tm details:Additives sucrose (1.5563 M),, pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: dHvH details:Additives sucrose (1.5563 M),, pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: Tm pH:2.8, details:Additives sucrose (1.0801 M),, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: dHvH pH:2.8, details:Additives sucrose (1.0801 M),, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: Tm pH:2.8, details:Additives sucrose (0.5632 M),, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: dHvH pH:2.8, details:Additives sucrose (0.5632 M),, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: Tm details:Additives sucrose (0 M),, pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: dHvH details:Additives sucrose (0 M),, pH:2.8, buffers:Glycine-HCl: 0.04 M ; Experimental Assay: Tm pH:4.2, details:Additives , buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:4.2, details:Additives , buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:4.2, details:Additives sucrose (1.9963 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:4.2, details:Additives sucrose (1.9963 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:4.2, details:Additives sucrose (1.5563 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:4.2, details:Additives sucrose (1.5563 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:4.2, details:Additives sucrose (1.0801 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:4.2, details:Additives sucrose (1.0801 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:4.2, details:Additives sucrose (0.5632 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:4.2, details:Additives sucrose (0.5632 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:4.2, details:Additives sucrose (0 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:4.2, details:Additives sucrose (0 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:5.5, details:Additives , buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:5.5, details:Additives , buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm details:Additives sucrose (1.9963 M),, pH:5.5, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH details:Additives sucrose (1.9963 M),, pH:5.5, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm details:Additives sucrose (1.5563 M),, pH:5.5, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH details:Additives sucrose (1.5563 M),, pH:5.5, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:5.5, details:Additives sucrose (1.0801 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:5.5, details:Additives sucrose (1.0801 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm pH:5.5, details:Additives sucrose (0.5632 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH pH:5.5, details:Additives sucrose (0.5632 M),, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: Tm details:Additives sucrose (0 M),, pH:5.5, buffers:Acetic acid-Sodium acetate: 0.04 M ; Experimental Assay: dHvH details:Additives sucrose (0 M),, pH:5.5, buffers:Acetic acid-Sodium acetate: 0.04 M
Libraries Mutations for sequence SKSKPDTSAPTSPKVTYKAPVPSGEVYFADSFDRGTLSGWILSKAKKDDTDDEIAKYDGKWEVDEMKETKLPGDKGLVLMSRAKHHAISAKLNKPFLFDTKPLIVQYEVNFQNGIECGGAYVKLLSKTPELNLDQFHDKTPYTIMFGPDKCGEDYKLHFIFRHKNPKTGVYEEKHAKRPDADLKTYFTDKKTHLYTLILNPDNSFEILVDQSIVNSGNLLNDMTPPVNPSREIEDPEDQKPEDWDERPKIPDPDAVKPDDWNEDAPAKIPDEEATKPDGWLDDEPEYVPDPDAEKPEDWDEDMDGEWEAPQIANPKCESAPGCGVWQRPMIDNPNYKGKWKPPMIDNPNYQGIWKPRKIPNPDFFEDLEPFKMTPFSAIGLELWSMTSDIFFDNFIVCGDRRVVDDWANDGWGLKKAADGAAEP ; Mutations for sequence KETAAAKFERQHMDSSTSAASSSNYCNQMMKSRNLTKDRCKPVNTFVHESLADVQAVCSQKNVACKNGQTNCYQSYSTMSITDCRETGSSKYPNCAYKTTQANKHIIVACEGNPYVPVHFDASV ; Mutations for sequence KVFGRCELAAAMKRHGLDNYRGYSLGNWVCAAKFESNFNTQATNRNTDGSTDYGILQINSRWWCNDGRTPGSRNLCNIPCSALLSSDITASVNCAKKIVSDGNGMNAWVAWRNRCKGTDVQAWIRGCRL
Sequence Assay Result Units