Pressure and Temperature Effects on Protein Solutions

1997 Fiscal Year Final Research Report Summary

• Project/Area Number: 08045025
• Research Category: Grant-in-Aid for international Scientific Research
• Allocation Type: Single-year Grants
• Section: University-to-University Cooperative Research
• Research Field: Physical chemistry
• Research Institution: Ritsumeikan University
• Principal Investigator: SAWAMURA Seiji Ritsumeikan Univ., Chem., Asso.Prof., 理工学部, 助教授 (10167439)
• Co-Investigator(Kenkyū-buntansha): ペィティ グレンフェル ブリティッシュコロンビア大学, 理学部, 教授 ; マクギリブレイ ロス ブリティッシュコロンビア大学, 理学部, 教授 ; マウク グラント ブリティッシュコロンビア大学, 医学部, 教授 ; KATO Minoru Ritsumeikan univ., Chem., Instructs, 理工学部, 講師 (00241258) ; TANIGUCHI Yoshihiro Ritsumeikan univ., Chem., Prof., 理工学部, 教授 (70066702) ; MAUK Grant Univ.of British Columbia, Biochem.Prof. ; MACGILLIVRAY Ross J.A. Univ.of British Columbia, Biochem.Prof. ; PATEY Grenfell N. Univ.of British Colimbia, Chem., Prof.
• Project Period (FY): 1996 – 1997
• Keywords: protein / pressure / Lysozyme

Research Abstract

Excess partial molar enthalpy of lysozyme in aqueous alcohol was measured using isothermal titration calorimetry. It was suggested that occupancy by the alcohol of a binding site on the surface of the protein may not be necessary for the alcohol to affect the properties of the protein. Pressure dependences of the solubiity for lysozyme and glycine in water measured. The formaer increased and the latter decreased with increasing pressure though both solubilities increased with increasing temperature. It suggests that the pressrue is one of key parameter to consider the protein hydration in the stand point of the constituent amino acids.
High-pressure FT-IR of Myoglobin (Mb) was measured in the range -10-70ﾟC.Pressure dependence of the second structure of Mb follows two state model. The structure irreversibly changes to random coil forming intermolecular beta-sheet with increasing temperature though it becomes reversible at low temperature.
It was, succeeded in expressing yeast cytochrome c in E.coli through construction of a plasmid for the co-expression of cytochrome c heme lyase and cytochrome c. The yield of wild-type cytochrome c from this system is relatively high. Variants in which the crucial Met80 ligand to the heme iron is replaced with Ala and that do no support respiration can be expressed in E.coli demonstrated that Lys72 is not trimethyllated postsynthetically as it is when expressed in yeast. This finding is no surprising because presumably E.coli does not posses the transmethylase enzyme that catalyzes this modification reaction.

Research Products

• [Publications] T.Nakai: "Effects of Pressure and Temperature on the viscosity β coefficient for Potassium Chloride in Water" Mutter.Sei.Res.Int.,. 3. 204-209 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] N.Takeda: "Pressure-inaucea structural rearrangemerts of bovine pancreatic trypsin inhibitor studied by FTIR spectroscopy" Biospectroscopy. (印刷中). (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] T.Matsumoto: "Effect of Pressure on the mechanism of hyarolysis of maltoteraose,maltopenthose,and maltohexose Catalyzei by porcine pancreatic α-amylase" Biochim,Bicphys,Acta,. 134・3. 243-250 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] K.Fumino: "NMR Studies on dynamic structure of hydrated water molecules in tetraalkylammonium bromide-D_2O solutions at 5-25℃" J.Mol.Liquids,. 72 (印刷中). (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] K.Fumino: "Concentration dependence of nuclear magnetic relaxation rates of alkalimetal ionic nucliein dilute solutions." Denki Kagaku. 65. 198-203 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Y.Kogh: "“Intermolecular Interactions"in Agueous Solutions of Three Components including Lysozyme" Fluid Phase Equil,. (印刷中). (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] T.J.F.Day: "Ion Solration in Binavy Mixtares" J.chem.Phys.106. 2782-2793 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] G.Ayton: "Liquid Crystal Phases of Dipolar Discotic Particles" Phys.Rer.E. 55. 447-454 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] M.Twitchett: "Intramolecular Electron Transfer Kinetics of a Synthetic Flavocytochrome C" J.Am.Chem.Soc.119. 435-436 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] J.Bujons: "Charge,Reversal of a Critical Actire Site Residue of Cytochrome C Peroxidase:Characterization of the AFS.48 Glu Variant" Eur.J.Biochem. 243. 72-84 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] C.L.Hunter: "Role of Heme Propionates in the Interaction of Heme with Apomyoglobin and Apocytochrome b_5." Biochemistry. 36. 1010-1017 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] C.L.Hunter: "Dissociation of Heme from Myoglobin and Cytochrome b_5:Comparison of Behaviorin Solution and the Gas Phase" Biochemistry. 36. 1018-1025 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] P.Westh: "Intermolecular Interactions of Lysozyme and Small Alcohols:A Calorimetric Investigation" J.Phys.Chem.B101. 5755-5758 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Y.Xu: "Light Scattering Study of Sol-Gel Transformation of SiO_2 Colloid at Extremely High Concentrations" Clays and Clay Minerals. (印刷中). (1998)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] D.R.Berard: "Structure of the Metal-Aqueous Electrolyte Solution Intertace" J.chem.phys.107. 4719-4728 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] J.C.Shelley: "Modeling and Structure of Mercury-Water Interfaces" J.chem.Phys.107. 2122-2141 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] G.Ayton: "Ferroelectric and Dipolar Glass Phases of Non-Crystalline Systems" Phys.Rev.E. 56. 562-570 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] N.M.Cann: "An Investigation of the Influence of Solute Size and Insertion on solvation Thermodynamics" J.Chem.Phys.106. 8165-8195 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] J.M.Nocek: "Cyclic Electron Transfer within the[Zn-deuteremyoylobin,Cytochrome b_5]Complex:Redox-Linked changes at the Interface" J.Am,chem,Loi.119. 2146-2155 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] A.G.Mank: "Control of Metalloprotein Redox Potentials:What Does Site-Directed Mutagenesis of Hemoproteins Tell Us?" J.Biol.Inorg.Chem.2. 119-125 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] K.Kishi: "Site-Directed Mutations of Phenylalanire-190 of Manganese Peroxidase:Effects on Stability,Function,and Coordination" Biochemistry. 36. 4268-4277 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] L.Konermann: "Acid-Induced Denaturation of Myoglobin Studied by Time-Resolved Electrospray Ionization Mass Spectrometry" Biochemistry. 36. 6448-6454 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] R.Maurns: "A Myoglobin Variant with a polar Substitution in Conserved Hydrophobic Cluster in the Heme Binding Pocket" Biochin.Biophys,Actu.,. 1341. 1-13 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Cote H.C.F: "A novel approach for the characterization and epitope determination of alupus anticoagulant-associated neutralizing anti-prothrombin antibody" Am.J.Clin.Putho.107. 197-205 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Cote,H.C.F.: "Functional characterization of recombinant human meizothrombin and meizothrombin(DesF1)" J.Biol.Chem.272. 6194-6200 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] M.A.Brown: "Vectors for expressing proteins at the amino-terminus of an activation domain for use in the yeast two-hybrid system" Anal.Biochem.247. 451-452 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Q-Y.He: "Effects of mutations of aspartic acid63 on the metal-binding properties of the recombinant N-lobe of human serum transferrin" Biochemistry. 36. 5522-5528 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] L.A.Luck: "19F NMR studies of the recombinant human transferrin N-lobe and three singlepoint mutants" Mag.Reson.Chem,. 35. 477-481 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] A.B.Mason: "Receptor recognition sites reside in both lobes of humann serun transfenin" Biochem.J.326. 77-85 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] O.Zak: "A kinetically active site in the C-lobe of human tramsferrin" Biochemistry. 36. 11036-11043 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] M.M.Guarna: "On-line monitoring and control of methanol concentration in shake-flask cultures of Pichia Pastoris" Biotechnol.Bioeng,. 56. 279-286 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Q-Y.He: "Inequivalence of the two tyrosine ligandsin the N-lobe of human serum transferrrin" Biochemistry. 36. 14853-14800 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] 谷口 吉弘: "生物・環境産業のための非加熱プロセス事典" サイエンスフォーラム, 95-99 (1997)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] T.Nakai, S.Sawamura, Y.Taniguchi, and T.Kuboyama: "Effects of Pressure and Temperature on the Viscosity B Coefficient for Potassium Chloride in Water" Matter.Sci.Res.Int.3. 204-209 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] N.Takeda, K.Nakano, M.Kato and Y.Taniguchi: "Pressure-induced Structural Rearrangements of Bovine Pancreatic Trypsin Inhibitor Studies by FTIR Spectroscopy" Biospectroscopy. (in press). (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] T.Matsumoto, S.Makimoto, and T.Taniguchi: "Effect of Pressure on the Mechanism of Hydrolysis of Maltoteraose, Maltopentaose, and Maltohexose Catalyzed by Porcine Pancreatic alpha-amylase" Biochim.Biophys.Acta. 1343. 243-250 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] K.Fumino, K.Yukiyasu, A.Shimizu, and Y.Taniguchi: "NMR Studies on Dynamic Structure of Hydrated Water Molecules in Tetraalkylammonium Bromide-D_2O solutions at 5-25ﾟC" J.Mol.Liquids. 72 (in press). (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] K.Fumino, A.Shimizu, and Y.Taniguchi: "Concentration Dependence of Nuclear Mgnetic Relaxation Rates of Alkali Metal Ionic Nuclie in Dilute Solutions" Denki Kagaku. 65. 198-203 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] K.Koga, P.Westh, Ch.Trandum, and C.A.Haynes: ""Intremolecular Interactions" in Aqueous Solutions of Three Components including Lysozyme" Fluid Phase Equil.(in press). (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Y.Taniguchi: "Property of Water and Aqueous Solution under High Pressure" Dictionary of Non-heating Process for Life-and Circumstances-Industry. 95-99 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] T.J.F.Day and G.N.Patey: "Ion Solvation in Binary Mixtures" J.Chem.Phys.106. 2782-2793 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] G.Ayton, D.Q.Wei.and G.N.Patey: "liquid Crystal Phases of Dipolar Discotic Particles" Phys.Rev.E.55. 447-454 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] M.Twitchett, J.C.Ferrer, P.Siddarth, and A.G.Mauk: "Intramolecular Electron Transfer Kinetics of a Synthetic Flavocytochrome c" J.Am.Chem.Soc.119. 435-436 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] J.Bujons, A.Dikiy, J.C.Ferrer, L.Banci, and A.G.Mauk: "Change Reversal of a Critical Active Site Residue of Cytochrome c Peroxidase : Characterization of the Arg48Glu Variant" Eur.J.Biochem.243. 72-84 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] C.L.Hunter, E.Lloyd, L.D.Eltis, S.P.Rafferty, H.Lee, M.Smith, and A.G.Mauk: "Role of Heme Propionates in the Interaction of Heme with Apomyoglobin and Apocytochrome b" Biochemistry. 36. 1010-1017 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] C.L.Hunter, A.G.Mauk, and D.J.Douglas: "Dissociation of Heme from Myoglobin and Cytochrome b_5 : Comparison of Behavior in Solution and the Gas Phase" Biochemistry. 36. 1018-1025 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] P.Westh and Y.Koga: "Intermolecular Interactions of Lysozyme and Small Alcohols : A Calorimetric Investigation" J.Phys.Chem.B. 101. 5755-5758 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Y.Xu and Y.Koga: "Light Scattering Study of Sol-Gel Transformation of SiO_2 Colloid at Extremely High Concentrations" Clays and Clay Minerals. (in press). (1998)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] D.R.Berard, M.Kinoshita, N.M.Cann, and G.N.Patey: "Structure of the Metal-Aqueous Elctrolyte Solution Interface" J.Chem.Phys.107. 4719-4728 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] J.C.Shelley, G.N.Patey, D.R.Berard, and G.M.Torrie: "Modeling and Structure of Mercury-Water Interfaces" J.Chem.Phys.107. 2122-2141 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] G.Ayton, M.J.P.Gingras, and G.N.Patey: "Ferroelectric and Dipolar Glass Phases of Non-Crystalline Systems" Phys.Rev.E. 56. 562-570 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] N.M.Cann and G.N.Patey: "An Investigation of the Influence of Solute Size and Insertion on Solvation Thermodynamics" J.chem.Phys.106. 8165-8195 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] J.M.Nocek, B.P.Sishta, J.C.Cameron, A.G.Mauk, and B.M.Hoffman: "Cyclic Electron Transfer within the [Zn-deuteromyoglobin, Cytochrome b_5] Complex : Redox-Linked Changes at the Interface" J.Am.Chem.Soc.119. 2146-2155 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] A.G.Mauk and G.R.Moore: "Control of Metalloprotein Redox Potentials : What Does Site-Directed Mutagenesis of Hemoproteins Tell Us?" J.Biol.Inorg.Chem.2. 119-125 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] K.Kishi, D.P.Hildebrand, M.Kusters-van Someren, J.Gettemy, A.G.Mauk, and M.H.Gold: "Site-Directed Mutations of Phenylalanin-190 of Manganese Peroxidase : Effects on stability, Function, and Coordination" Biochemistry. 36. 4268-4277 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] L.Konermann, F.I.Rosell, A.G.Mauk, and D.J.Douglas: "Acid-Induced Denaturation of Myoglobin Studies by Time-Resolved Electrospray Ionization Mass Spectrometry" Biochemistry. 36. 6448-6454 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] R.Maurus, C.M.Overall, R.Bogunil, Y.Luo, A.G.Mauk, M.Smith, and G.D.Brayr: "A Myoglobin Variant with a Polar Substitution in a Conserved Hydrophobic Cluster in the Heme Binding Pocket" Biochem.Biophys.Acta. 1341. 1-13 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] H.C.F.Cote, D.G.Huntsman, J.Wu, L.D.Wadsworth, and R.T.A.MacGillivray: "A Novel Approach for the Characterization and Epitope Determination of a Lupus Anticoagulant-associated Neutralizing Anti-prothrombin Antibody" Am.J.Clin.Pathol.107. 197-205 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] H.C.F.Cote, L.Bajzar, W.K.Stevens, J.A.Samis, J.Morser, R.T.A.MacGillivray, and M.E.Nesheim: "Functional Characterization of Recombinant Human Meizothrombin and Meizothrombin (DesF1). Thrombomodulin-dependent Activation of Protein C and Thrombin-activatable Fibrinolysis Inhibitor (TAFI), Platelet Aggregation, Antithrombin-III Inhibition" J.Biol.Chem.272. 6194-6200 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] M.A.Brown and R.T.A.MacGillivray: "Vectors for Expressing Proteins at the Amino-terminus of an Activation Domain for Use in the Yeast Two-hybrid System" Anal.Biochem.247. 451-452 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Q.-Y.He, A.B.Mason, R.C.Woodworth, B.M.Tam, T.Wadsworth, and R.T.A.MacGillivray: "Effects of Mutations of Aspartic Acid 63 on the Metalbinding Properties of the Recombinant N-lobe of Human Serum Transferrin" Biochemistry. 36. 5522-5528 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] L.A.Luck, A.B.Mason, K.J.Savage, R.T.A.MacGillivray, and R.C.Woodworth: "19F NMR Studies of the Recombinant Human Transferrin N-lobe and Three Single Point Mutants" Mag.Reson.Chem.35. 477-481 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] A.B.Mason, R.C.Woodworth, R.W.A.Oliver, B.N.Green, L.-N.Lin, J.F.Brandts, B.M.Tam, K.J.Savage, J.A.Lineback, and R.T.A.MacGillivray: "Receptor Recognition Sites Reside in Both Lobes of Human Serum Transferrin" Biochem.J.326. 77-85 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] O.Zak, B.Tam, R.T.A.MacGillivray, and P.Aisen: "A Kinetically Active Site in the C-lobe of Human Transferrin" Biochemistry. 36. 11036-11043 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] M.M.Guama, G.L.Lesnicki, B.Tam.J.Robinson, C.Z.Radziminski, D.Haseniwinkle, A.Boraston, E.Jervis, R.T.A.MacGillivray, R.F.B.Turner, and D.G.Kilburn: "On-line Monitoring and Control of Methanol Concentration in Shake-flask Cultures of Pichia pastoris" Biotechnol.Bioeng.56. 279-286 (1997)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Q.-Y.He, A.B.Mason, R.C.Woodworth, B.M.Tam, R.T.A.MacGillivray, J.K.Grady, and N.D.Chasteen: "Inequivalence of the Two Tyrosine Ligands in the N-lobe of Human Serum transferrin" Biochemistry. 36. 14853-14860 (1997)
  • Description: 「研究成果報告書概要(欧文)」より