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TitleTitle SUPRAMOLECULAR MODELS OF SOLID PROTEINS V.V. Gorbatchuk, M.A.Ziganshin, N.A.Mironov, I.S.Antipin, B.N.Solomonov, A.I.Konovalov Department of.

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Presentation on theme: "TitleTitle SUPRAMOLECULAR MODELS OF SOLID PROTEINS V.V. Gorbatchuk, M.A.Ziganshin, N.A.Mironov, I.S.Antipin, B.N.Solomonov, A.I.Konovalov Department of."— Presentation transcript:

1 TitleTitle SUPRAMOLECULAR MODELS OF SOLID PROTEINS V.V. Gorbatchuk, M.A.Ziganshin, N.A.Mironov, I.S.Antipin, B.N.Solomonov, A.I.Konovalov Department of Chemistry, Kazan State University, Kazan, Russia

2 Problems: - Thermodynamic description of the role of water in the protein-substrate binding - Thermodynamic criteria of biocompatibility Strategy: Comparison of the structure - binding property relationships for proteins and more simple objects

3 Choice of protein models Solutions Crystalline organic hosts Proteins Structure Property Polymers Zeolites

4 Object substrate vapor solid protein + S S Protein H2OH2O H2OH2O H2OH2O H2OH2O H2OH2O H2OH2O + n H 2 O Receptor properties of enzymes and antibodies in water solutions and in solid state are similar A.M.Klibanov, Trends Biotechnol. (2000) 18, 85enzymes S.S.Setford, Trends Anal. Chem. (2000) 19, antibodies Effect of substrate dehydration masks the molecular recognition S Enzymatic reactions in solid phase Khurgin Yu. I., Maksareva E. Yu, Bioorg. Khim 17(1991) Gas-phase enzymatic reactors Yang F. and Russell A. J., Biotechnol. Bioeng., 49 (1996) 709. Enzymatic reactions in nonaqueous solvents P.J. Halling, Enzyme Microb. Technol. 16 (1994) Applications

5 Objects n-Nonane C6H6C6H6 C 6 H 5 OCH 3 n-PrOH CH 3 OHCHCl 3 c-Hexane CH 3 CN1,4-Dioxane n-Hexane n-Heptane EtOHCH 3 NO 2 Pinacolone o-Xylene EtCN PrCN Toluene C 6 H 5 Et (CH 3 ) 2 CO n-Octane CCl 4 Sorbates/substrates/guests t-BuOAc Pyridine 2,2’-(9-hydroxy-9- fluorenyl)biphenyl tert-butylthiacalix[4]arene Casein Albumin Invertase Trypsin Collagen  -Lactoglobulin  cyclodextrin poly(acrylic acid 6-aminohexylamide) n

6 Electropneumatic dosing B. Kolb, P. Pospisil, T. Borath and M. Auer, J. High Res. Chromatogr. & Chromatogr. Commun., 1979, 2, 283. Activity determination A. Hussam and P. W. Carr, Anal. Chem., 1985, 57, 793. Data examples J. H. Park, A. Hussam, P. Couasnon, D. Fritz and P. W. Carr, Anal. Chem., 1987, 59, Electropneumatic dosing B. Kolb, P. Pospisil, T. Borath and M. Auer, J. High Res. Chromatogr. & Chromatogr. Commun., 1979, 2, 283. Activity determination A. Hussam and P. W. Carr, Anal. Chem., 1985, 57, 793. Data examples J. H. Park, A. Hussam, P. Couasnon, D. Fritz and P. W. Carr, Anal. Chem., 1987, 59, ELECTROPNEUMATIC VALVE SPLITTER FUSED SILICA COLUMN VIAL WITH SAMPLE THERMOSTATE DEVICE OF STATIC HEADSPACE GAS CHROMATOGRAPHIC ANALYSIS solid phase binding of vapors in comparable conditions

7 Protein Isotherms Dried proteins hydration in g H2O/g protein high binding low binding More selective than zeolites V.V.Gorbatchuk, M.A.Ziganshin, B.N.Solomonov, M.D.Borisover, J.Phys.Org.Chem.,10 (1997) albumin Casein (0.06 h) Pure protein: V.V.Gorbatchuk, M.A.Ziganshin, N.A.Mironov, B.N.Solomonov, Biophys. Biochim. Acta. 1545(2001) Trypsin (0.05 h) Effect of lactose (~70%) Conclusion: solid proteins are nonporous objects

8 Contents of lactose is ~70% No binding without protein V.V.Gorbatchuk, M.A.Ziganshin, N.A.Mironov, B.N.Solomonov, Biophys. Biochim. Acta. 1545(2001) Trypsin (0.05 h) V. V. Gorbatchuk, A. G. Tsifarkin, I. S. Antipin, B. N. Solomonov, A. I. Konovalov, J. Seidel, F. Baitalov, J. Chem.Soc. Perkin Trans. 2, 2000, N11, P C6H6 V. V. Gorbatchuk, A. G. Tsifarkin, I. S. Antipin, B. N. Solomonov, A. I. Konovalov, P.Lhotak, I.Stibor J. Phys. Chem.B., 2002, 106(23); Phase transitionPhase transition clathrate formation & phase transition

9 Cooperativ e hydration effects protein hydration rate of enzymatic reaction Enthalpies of protein immersion in water- organic mixtures V.A.Sirotkin, M.D. Borisover, B.N.Solomonov, Biophys. Chem. 69 (1997) Gas-phase reactors: enzymatic reactions Gas-phase reactors: enzymatic reactions Yang F. and Russell A. J., Biotechnol. Bioeng., 49 (1996) 709. Enzymatic reactions in solid phase Khurgin Yu. I., Maksareva E. Yu, Bioorg. Khim 17(1991) Protein suspensions in nonaqueous solvents: -water sorption -enzymatic reactions Protein suspensions in nonaqueous solvents: -water sorption -enzymatic reactions P.J. Halling, Enzyme Microb. Technol. 16 (1994) Solid proteins in low-water conditions What is the hydration effect on the initial state?

10 Cooperative hydration effect Hydration threshold V.V. Gorbatchuk, M.A. Ziganshin, B.N. Solomonov, Biophys. Chem. 81 (1999) protein water substrate clathrates are formed ---- this process defines the activity of enzymes and binding ability of antibodies

11 n h,gH2O/g polymer or HSA Hydrophobic hydration EtOH Effect of hydrophobic hydration is a sufficient thermodynamic evidence of biocompatibility

12 Higher temperature is necessary for protein to function in low-water conditions high effect for hydrophobic compounds No effect for hydrophilic compound G. Brausse et al. JACS V.72. P opposite effect for water  -Lactoglobulin

13 Cooperation with Dr. W.Habicher TU Dresden, Germany Dr. Th.Haertle LEIMA INRA, Nantes, France Dr. M.D.Borisover Inst. Soil & Water, Bet Dagan, Israel Acknowledgements Participants: Dr. Ziganshin M.A. Dr. Tsifarkin A.G. Mironov N.A. Prof. Antipin I.S. Prof. Solomonov B.N. Prof. Konovalov A.I. Grants URBR ( ), BRHE (REC-007) ANT ( )


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