本研究を行うに際し、終始御指導、御鞭撻を賜りました熊本大学大学院医学 薬学研究部・薬物動態制御学分野、小田切優樹教授に深甚なる感謝の意を表し ます。
本研究を行うにあたり、終始有益な御助言と御便宜を惜しまれなかった Cincinnati大学H. Brian Halsall教授に心より感謝致します。
本論文作成に際し、有益な御教示と御校閲を賜りました熊本大学大学院医学 薬学研究部・生命分析化学分野、宇野公之教授、機能分子構造解析学分野、山 縣ゆり子教授ならびに製剤設計学分野、平山文俊助教授に心から感謝致します。
本研究に際し、終始有益な御助言と御便宜を惜しまれなかった熊本大学大学 院医学薬学研究部・薬物動態制御学分野、末永綾香助手ならびに丸山 徹博士 に心より感謝致します。
本研究に際し、有益な御助言と御協力を頂きました坂井紀文修士(三菱化学 安全科学研究所)ならびに小嶺嘉男修士(長崎大学医学部附属病院薬剤部)に 深謝致します。
リポソーム作製に際し、御指導、御協力頂きました京都大学大学院薬学研究 科、半田哲郎教授に深謝致します。
AGPのcDNAを恵与して頂いた協和発酵工業株式会社に心より御礼申し上げ ます。
AGP のポリクローナル抗体作製に際し、御指導、御協力頂きました熊本大学 大学院薬学教育部・細胞機能分子解析学研究室、清水英介修士に心から感謝致 します。
本研究を行うに際し、御協力頂いた福永直子学士をはじめ、熊本大学薬学部・
薬物動態制御学研究室の諸氏に心から感謝致します。
最後にここまで私を支えてくれた両親ならびに妻、裕美に心から感謝致しま す。
平成16年 3月
参考文献
1) H.B. Halsall, R.C. Austin, J.L. Dage, H. Sun, K.T. Schlueter, In:Proc. int. symp.
On serum albumin and alpha1-acid glycoprotein (Eds. M Otagiri, Y Sugiyama, B Testa, J-P Tillement), Tokyo Print, Kumamoto, Japan, pp45, (2000).
2) J.P. Aubert, M.H. Loucheux-Lefebvre, Conformational study of alpha1-acid glycoprotein., Arch. Biochem. Biophys., 175, 400-409 (1976).
3) A. Rojo-Dominguez, A. Hernandez-Arana, Three-dimensional modeling of the protein moiety of human alpha1-acid glycoprotein, a lipocalin-family member., Protein Seq. Data Anal., 5, 349-355 (1993).
4) S. Pervaiz and K. Brew, Homology and structure-function correlations between alpha1-acid glycoprotein and serum retinol-binding protein and its relatives., FASEB J., 1, 209-214 (1987).
5) J. Pevsner, R.R. Reed, P.G. Feinstein, S.H. Snyder, Molecular cloning of odorant-binding protein: member of a ligand carrier family., Science, 241, 336-339 (1988).
6) A.J. Clark, P.M. Clissold, R.A. Shawi, P. Beattie, J. Bishop, Structure of mouse major urinary protein genes: different splicing configurations in the 3'-non-coding region., EMBO J., 3, 1045-1052 (1984).
7) J.M.H. Kremer, J. Wilting, L.H.M. Janssen, Drug binding to human alpha-1-acid glycoprotein in health and disease. [Review], Pharmacol. Rev., 40, 1-47 (1988).
8) H. Bacchus, Serum seromucoid and acid mucopolysaccharide in malignant neoplastic diseases., Cancer, 18, 1285-1291 (1965).
9) J.A. Martyn, D.R. Abernethy, D.J. Greenblatt, Plasma protein binding of drugs after severe burn injury., Clin. Pharmacol. Ther., 35, 535-539 (1984).
10) O. Bertaux, T. Fournier, L. Chauvelot-Moachon, D. Porquet, R. Valencia, G.
Durand, Modifications of hepatic alpha-1-acid glycoprotein and albumin gene expression in rats treated with phenobarbital., Eur. J. Biochem., 203, 655-661 (1992).
11) L. Chauvelot-Moachon, F. Delers, C. Pous, R. Engler, F. Tallet, J.P. Giroud, Alpha1-acid glycoprotein concentrations and protein binding of propranolol in Sprague-Dawley and Dark Agouti rat strains treated by phenobarbital., J.
Pharmacol. Exp. Ther., 244, 1103-1108 (1988).
12) J.W. Holladay, M.J. Dewey, S.D. Yoo, Steady-state kinetics of imipramine in transgenic mice with elevated serum AAG levels., Pharm. Res., 13, 1313-1316 (1996).
13) N. Kurata, S. Matsushita, K. Nishi, H. Watanabe, S. Kobayashi, A. Suenaga, M.
Otagiri, Characterization of a binding site of UCN-01, a novel anticancer drug on alpha-acid glycoprotein., Biol. Pharm. Bull., 23, 893-895 (2000).
14) T.H. Lin, Y. Sawada, Y. Sugiyama, T. Iga, M. Hanano, Effects of albumin and alpha1-acid glycoprotein on the transport of imipramine and desipramine through the blood-brain barrier in rats., Chem. Pharm. Bull., 35, 294-301 (1987).
15) R. Weisiger, J. Gollan, R. Ockner, Receptor for albumin on the liver cell surface may mediate uptake of fatty acids and other albumin-bound substances., Science, 211, 1048-1051 (1981).
16) E.L. Forker, B.A. Luxon, Albumin helps mediate removal of taurocholate by rat liver., J. Clin. Invest., 67, 1517-1522 (1981).
17) E.L. Forker, B.A. Luxon, Albumin-mediated transport of rose bengal by perfused rat liver. Kinetics of the reaction at the cell surface., J. Clin. Invest., 72, 1764-1771 (1983).
18) M.M. Andersen, Leucocyte-associated plasma proteins. Association of prealbumin, albumin, orosomucoid, alpha1-antitrypsin, transferrin and haptoglobin with human lymphocytes, monocytes, granulocytes and a promyelocytic leukaemic cell line (HL-60)., Scand. J. Clin. Lab. Invest., 43, 49-59 (1983).
19) H. Maeda, T. Morinaga, I. Mori, K. Nishi, Further characterization of the effects of alpha1-acid glycoprotein on the passage of human erythrocytes through micropores., Cell Struct. Funct., 9, 279-290 (1984).
20) D. Predescu, S. Predescu, T. McQuistan, G.E. Palade, Transcytosis of alpha1-acidic glycoprotein in the continuous microvascular endothelium., Proc. Natl.
Acad. Sci. U.S.A., 95, 6175-6180 (1998).
21) T. Horie, T. Mizuma, S. Kasai, S. Awazu, Conformational change in plasma albumin due to interaction with isolated rat hepatocyte., Am. J. Physiol., 254, G465-G470 (1988).
22) V.E. Bychkova, R.H. Pain, O.B. Ptitsyn, The 'molten globule' state is involved in the translocation of proteins across membranes?., FEBS Lett., 238, 231-234 (1988).
23) V.E. Bychkova, O.B. Ptitsyn, The functional state of denatured proteins: the principles of modeling and the first results. [Review], Tsitologiia, 37, 1238-1250 (1995).
24) F.G. van der Goot, J.M. Gonzales-Manas, J.H. Lakey, F. Pattus, A 'molten-globule' membrane-insertion intermediate of the pore-forming domain of colicin A., Nature, 354, 408-410 (1991).
25) M. Hase, T. Yoshimi, Y. Ishikawa, A. Ohba, L. Guo, S. Mima, T. Mizushima, Site-directed mutational analysis for the membrane binding of DnaA protein.
Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids., J. Biol. Chem., 273, 28651-28656 (1998).
26) M. Makise, S. Mima, T. Tsuchiya, T. Mizushima, Identification of amino acids involved in the functional interaction between DnaA protein and acidic phospholipids., J. Biol. Chem., 275, 4513-4518 (2000).
27) J.K. Davies, E.A. Thumser, D.C. Wilton, Binding of recombinant rat liver fatty acid-binding protein to small anionic phospholipid vesicles results in ligand release: a model for interfacial binding and fatty acid targeting., Biochemistry, 38, 16932-16940 (1999).
28) E.R. Smith, J. Storch, The adipocyte fatty acid-binding protein binds to membranes by electrostatic interactions., J. Biol. Chem., 274, 35325-35330 (1999).
29) N. Arispe, E. Rojas, H.B. Pollard, Alzheimer disease amyloid beta protein forms calcium channels in bilayer membranes: blockade by tromethamine and aluminum., Proc. Natl. Acad. Sci. U.S.A., 90, 567-571 (1993).
30) M. Morillas, W. Swietnicki, P. Gambetti, W.K. Surewicz, Membrane environment alters the conformational structure of the recombinant human prion protein., J.
Biol. Chem., 274, 36859-36865 (1999).
31) P.L. Luisi, M. Giomini, M.P. Pileni, B.H. Robinson, Reverse micelles as hosts for proteins and small molecules. [Review], Biochim. Biophys. Acta, 947, 209-246 (1988).
32) K. Martinek, A.V. Levashov, N. Klyachko, Y.L. Khmelnitski, I.V. Berezin, Micellar enzymology. [Review], Eur. J. Biochem., 155, 453-468 (1986).
33) T. Maruyama, M. Otagiri, A. Takadate, Characterization of drug binding sites on alpha1-acid glycoprotein., Chem. Pharm. Bull., 38, 1688-1691 (1990).
34) T. Miyoshi, K. Sukimoto, M. Otagiri, Investigation of the interaction mode of phenothiazine neuroleptics with alpha1-acid glycoprotein., J. Pharm. Pharmacol., 44, 28-33 (1992).
35) H. Wojtasek, W.S. Leal, Conformational change in the pheromone-binding protein from Bombyx mori induced by pH and by interaction with membranes., J. Biol.
Chem., 274, 30950-30956 (1999).
36) E. de Leeuw, K. te Kaat, C. Moser, G. Menestrina, R. Demel, B. de Kruijff, B.
Oudega, J. Luirink, I. Sinning, Anionic phospholipids are involved in membrane association of FtsY and stimulate its GTPase activity., EMBO J., 19, 531-541 (2000).
37) R. Halaban, E. Cheng, S. Svedine, R. Aron, D.N. Hebert, Proper folding and endoplasmic reticulum to golgi transport of tyrosinase are induced by its substrates, DOPA and tyrosine., J. Biol. Chem., 13, 11933-11938 (2000).
38) K.F. Winklhofer, U. Heller, A. Reintjes, J. Tatzelt, Inhibition of complex glycosylation increases the formation of PrPsc., Traffic, 4, 313-322 (2003).
39) J. Zhu, I. Watanabe, A. Poholek, M. Koss, B. Gomez, C. Yan, E. Recio-Pinto, W.B. Thornhill, Allowed N-glycosylation sites on the Kv1.2 potassium channel S1-S2 linker: implications for linker secondary structure and the glycosylation effect on channel function., Biochem. J., 375, 769-775 (2003).
40) K.S. Kwon , M.H. Yu, Effect of glycosylation on the stability of alpha1-antitrypsin toward urea denaturation and thermal deactivation., Biochim. Biophys. Acta, 1335, 265-272 (1997).
41) K. Yang, A. Basu, M. Wang, R. Chintala, MC. Hsieh, S. Liu, J. Hua, Z. Zhang, J.
Zhou, M. Li, H. Phyu, G. Petti, M. Mendez, H. Janjua, P. Peng, C. Longley, V.
Borowski, M. Mehlig, D. Filpula, Tailoring structure-function and pharmacokinetic properties of single-chain Fv proteins by site-specific PEGylation., Protein Eng., 16, 761-770 (2003).
42) A. Saito, M. Usui, Y. Song, H. Azakami, A. Kato, Secretion of glycosylated alpha-lactalbumin in yeast Pichia pastoris., J. Biochem., 132, 77-82 (2002).
43) H. Watanabe, K. Yamasaki, U. Kragh-Hansen, S. Tanase, K. Harada, A. Suenaga, M. Otagiri, In vitro and in vivo properties of recombinant human serum albumin from Pichia pastoris purified by a method of short processing time., Pharm. Res., 18, 1775-1781 (2002).
44) C. Ying, G. Boyu, S. Daxin, Y. Hanying, C. Yongqin, L. Yuyang, L. Xiang, T.
Peikun, G. Jianren, Expression of transthyretin gene in Pichia pastoris., Chin. J.
Biotechnol., 15, 211-217 (1999).
45) T.R. Kim, Y. Goto, N. Hirota, K. Kuwata, H. Denton, S.Y. Wu, L. Sawyer, C.A.
Batt, High-level expression of bovine beta-lactoglobulin in Pichia pastoris and characterization of its physical properties., Protein Eng., 10, 1339-1345 (1997).
46) A. Miyajima, K. Otsu, J. Schreurs, M.W. Bond, J.S. Abrams, K. Arai, Expression of murine and human granulocyte-macrophage colony-stimulating factors in S.
cerevisiae: mutagenesis of the potential glycosylation sites., EMBO J., 5, 1193-1197 (1986).
47) J.R. Albani, Motions of tryptophan residues in asialylated human alpha1-acid glycoprotein., Biochim. Biophys. Acta, 1291, 215-220 (1996).
48) Y.H. Chen, J.T. Yang, H.M. Martinez, Determination of the secondary structures of proteins by circular dichroism and optical rotatory dispersion., Biochemistry, 11, 4120-4131 (1972).
49) R.D. King, M. Saqi, R. Sayle, M.J. Sternberg, DSC: public domain protein secondary structure predication., Comput. Appl. Biosci., 13, 473-474 (1997).
50) Y. Guermeur, C. Geourjon, P. Gallinari, G. Deleage, Improved performance in protein secondary structure prediction by inhomogeneous score combination., Bioinformatics, 15, 413-421 (1999).
51) B, Rost, C. Sander, R. Schneider, PHD: an automatic mail server for protein secondary structure prediction., Comput. Appl. Biosci., 10, 53-60 (1994).
52) F. Eisenhaber, C. Frommel, P. Argos, Prediction of secondary structural content of proteins from their amino acid composition alone. II. The paradox with secondary structural class., Proteins, 25, 169-179 (1996).
53) C. Geourjon, G. Deleage, SOPMA: significant improvements in protein secondary structure prediction by consensus prediction from multiple alignments., Comput.
Appl. Biosci., 11, 681-684 (1995).
54) C. Geourjon, G. Deleage, SOPM: a self-optimized method for protein secondary structure prediction., Protein Eng., 7, 157-164 (1994).
55) G. Deleage, B. Roux, An algorithm for protein secondary structure prediction based on class prediction., Protein Eng., 1, 289-294 (1987).
56) C. Tanford, Protein denaturation. [Review], Adv. Protein Chem., 23, 121-282 (1968).
57) P.D. Thomas, K.A. Dill, Local and nonlocal interactions in globular proteins and mechanisms of alcohol denaturation., Protein Sci., 2, 2050-2065 (1993).
58) F.D. Sonnichsen, J.E. Van Eyk, R.S. Hodres, B.D. Sykes, Effect of trifluoroethanol on protein secondary structure: an NMR and CD study using a synthetic actin peptide., Biochemistry, 31, 8790-8798 (1992).
59) K. Shiraki, K. Nishikawa, Y. Goto, Trifluoroethanol-induced stabilization of the alpha-helical structure of beta-lactoglobulin: implication for non-hierarchical protein folding., J. Mol. Biol., 235, 180-194 (1995).
60) D. Hamada, S. Segawa, Y. Goto, Non-native alpha-helical intermediate in the refolding of beta-lactoglobulin, a predominantly beta-sheet protein., Nature Struct.
Biol., 3, 868-873 (1996).
61) H.J. Dyson, G. Merutka, J.P. Waltho, R.A. Lerner, P.E. Wright, Folding of peptide fragments comprising the complete sequence of proteins., J, Mol, Biol., 226, 795-817 (1992).
62) J.J. Yang, M. Buck, M. Pitkeathly, M. Kotik, D.T. Havnie, C.M. Dobson, S.E.
Radford, Conformational properties of four peptides spanning the sequence of hen lysozyme., J. Mol. Biol., 252, 483-491 (1995).
63) Y. Kuroda, D. Hamada, T. Tanaka, Y. Goto, High helicity of peptide fragments corresponding to beta-strand regions of beta-lactoglobulin observed by 2D-NMR spectroscopy., Fold. Des., 1, 255-263 (1996).
64) J.W. Nelson, N.R. Kallenbach, Stabilization of the ribonuclease S-peptide alpha-helix by trifluoroethanol., Proteins, 1, 211-217 (1986).
65) N. Hirota, K. Mizuno, Y. Goto, Group additive contributions to the alcohol-induced alpha-helix formation of melittin: implication for the mechanism of the alcohol effects on proteins., J. Mol. Biol., 275, 365-378 (1998).
66) Y. Goto, A.L. Fink, Conformational states of beta-lactamase: molten-globule states at acidic and alkaline pH with high salt., Biochemistry, 28, 945-952 (1989).
67) Y. Goto, L.J. Calciano, A.L. Fink, Acid-induced folding of proteins., Proc. Natl.
Acad. Sci. U.S.A., 87, 573-577 (1990).
68) Y. Goto, A.L. Fink, Phase diagram for acidic conformational states of apomyoglobin., J. Mol. Biol., 214, 803-805 (1990).
69) V.E. Bychkova, A.E. Dujsekina, S.I. Klenin, E.I. Tiktopulo, V.N. Uversky, O.B.
Ptitsyn, Molten globule-like state of cytochrome c under conditions simulating those near the membrane surface., Biochemistry, 35, 6058-6063 (1996).
70) M.l. Friedman, K.T. Schlueter, T.L. Kirley, H.B. Halsall, Fluorescence quenching of human orosomucoid. Accessibility to drugs and small quenching agents., Biochem. J., 232, 863-867 (1985).
71) M.R. Eftink, Fluorescence techniques for studying protein structure. [Review], Methods Biochem. Anal., 35, 127-205 (1991).
72) O.B. Ptitsyn, in Protein Folding (Creighton, T. E., Ed), Freeman, New York, pp243 (1992).
73) A.I. Ivanov, V.B. Gavrilov, D.A. Furmanchuk, O.V. Aleinikova, S.V. Konev, G.V.
Kaler, Fluorescent probing of the ligand-binding ability of blood plasma in the acute-phase response., Clin. Exp. Med., 2, 147-155 (2002).
74) H. Imamura, T. Maruyama, H. Okabe, H. Shimada, M. Otagiri, A simple and rapid fluorometric determination method of alpha 1-acid glycoprotein in serum using quinaldine red., Pharm. Res., 11, 566-570 (1994).
75) H. Imamura, T. Maruyama, M. Otagiri, Evaluation of quinaldine red as a fluorescent probe for studies of drug-alpha1-acid glycoprotein interaction., Biol.
Pharm. Bull., 16, 926-929 (1993).
76) J.J. Ewbank, T.E. Creighton, Structural characterization of the disulfide folding intermediates of bovine alpha-lactalbumin., Biochemistry, 32, 3694-3707 (1993).
77) J.J. Ewbank, T.E. Creighton, Pathway of disulfide-coupled unfolding and refolding of bovine alpha-lactalbumin., Biochemistry, 32, 3677-3693 (1993).
78) M. Ikeguchi, S. Sugai, M. Fujino, T. Sugawara, K. Kuwajima, Contribution of the 6-120 disulfide bond of alpha-lactalbumin to the stabilities of its native and molten globule states., Biochemistry, 31, 12695-12700 (1992).
79) M. Yagi, K. Sakurai, C. Kalidas, C.A. Batt, Y. Goto, Reversible unfolding of bovine beta-lactoglobulin mutants without a free thiol group., J. Biol. Chem., 278, 47009-47015 (2003).
80) T. Croguennec, S. Bouhallab, D. Molle, B.T. O'Kennedy, R. Mehra, Stable monomeric intermediate with exposed Cys-119 is formed during heat denaturation of beta-lactoglobulin., Biochem. Biophys. Res. Commun., 301, 465-471 (2003).
81) V. Forge, M. Hoshino, K. Kuwata, M. Arai, K. Kuwajima, C.A. Batt, Y. Goto, Is folding of lactoglobulin non-hierarchic? Intermediate with native-like beta-sheet and non-native alpha-helix., J. Mol. Biol., 296, 1039-1051 (2000).
82) T.V. Burova, Y. Choiset, V. Tran, T. Haertle, Role of free Cys121 in stabilization of bovine beta-lactoglobulin B., Protein Eng., 11, 1065-1073 (1998).
83) V.P. Saxena, D.B. Wetlaufer, Formation of three-dimensional structure in proteins.
I. Rapid nonenzymic reactivation of reduced lysozyme., Biochemistry, 9, 5015-5023 (1970).
84) P.A. Jennings, P.E. Wright, Formation of a molten globule intermediate early in the kinetic folding pathway of apomyoglobin., Science, 262, 892-896 (1993).
85) D. Barrick, F.M. Hughton, R.L. Baldwin, Molecular mechanisms of acid denaturation. The role of histidine residues in the partial unfolding of apomyoglobin., J. Mol. Biol., 237, 588-601 (1994).
86) D. Barrick, Replacement of the proximal ligand of sperm whale myoglobin with free imidazole in the mutant His-93-->Gly., Biochemistry, 33, 6546-6554 (1994).
87) A. Rojo-Dominguez, R. Zubillaga-Luna, A. Hernandez-Arana, Unfolding behavior of human alpha1-acid glycoprotein is compatible with a loosely folded region in its polypeptide chain., Biochemistry, 29, 8689-8695 (1990).
88) M.Ganguly, R.H. Carnighan, U. Westphal, Steroid-protein interactions. XIV.
Interaction between human alpha1-acid glycoprotein and progesterone., Biochemistry, 6, 2803-2814 (1967).
89) P.L. Luisi, B. Steinmann-Hofmann, Activity and conformation of enzymes in reverse micellar solutions., Methods Enzymol., 136, 188-216 (1987).
90) N. A. Attallah, G. F. Lata, Steroid-protein interactions studies by fluorescence quenching., Biochim. Biophys. Acta, 168, 321-333 (1968).
91) J.R. Lakowicz, Principle of Fluorescence Spectroscopy, Plenum Press, New York, (1983).
92) 奥 直人, リポソームの作成と実験法, p36, 廣川書店, 東京 (1993).
93) New, R. R. C., in Liposomes : A Practical Approach (New, R. R. C., Ed).
pp33,Oxford University Press, Oxford, U. K. (1990).
94) C.A. Buser, C.T. Sigal, M.D. Resh, S. McLaughlin, Membrane Binding of Myristylated Peptides Corresponding to the NH2 Terminus of Src., Biochemistry, 33, 13093-13101 (1994).
95) M.M. Bradford, A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding., Anal. Biochem., 72, 248-54 (1976).
96) J. Braman, C. Papworth, A. Greener, Site-directed mutagenesis using double-stranded plasmid DNA templates. [Review], Methods Mol. Biol., 57, 31-44 (1996).
97) J.A. Oka, P.H. Weigel, Recycling of the asialoglycoprotein receptor in isolated rat hepatocytes. Dissociation of internalized ligand from receptor occurs in two kinetically and thermally distinguishable compartments., J. Biol. Chem., 258, 10253-10262 (1983).
98) L. Warren, The thiobarbituric acid assay of sialic acids., J. Biol. Chem., 234, 1971-1975 (1959).