Galectin-1蛋白是一種b-半乳糖苷結合蛋白，其參與細胞貼附、分化及生長調控。Gal-1具有6個還原態的硫氫基，此官能基和蛋白質之lectin活性有直接的關係，但是Gal-1在形成二聚體的過程，並無硫氫基的參與。由近年的研究指出，氧化態Gal-1喪失其lectin活性，但在低濃度下，可促使神經細胞再生及使細胞轉化。由實驗室初步結果顯示，As3+可和Gal-1上的硫氫基結合，並且造成Gal-1的氧化，而此種氧化態Gal-1，運用其生物性功能來影響CHOA細胞；而轉殖Gal-1基因的哺乳類動物細胞，會累積且吸收較多的砷化物。我們利用大腸桿菌系統純化Gal-1和Gal-As3+；並以CD觀察蛋白質，發現gal-As3+在200~205 nm之間有一負值訊號，而gal-1則在205~210nm之間有一負值訊號。由CD值位移的結果間接顯示，亞砷酸鈉 (As3+)和gal-1結合，導致蛋白質二級結構的改變。而由恆溫滴定微卡計的試驗結果得知，亞砷酸鈉滴入gal-1蛋白溶液中的吸附焓介於-55~-69 (kJ/mole As3+)之間。顯示亞砷酸鈉和gal-1之間的作用能量已超過配位鍵的範圍 (~21 kJ/mole)。由初步的結果得知，gal-As3+失去部分lectin活性，且影響HFW細胞的存活率。

Galectin-1 (gal-1), a b-galactoside binding protein, functions in cell adhesion, development, and growth regulation. Gal-1 has six free sulfhydryl groups, which are essential for their lectin property and do not involve in the formation of dimeric gal-1. Recent studies suggest that oxidized gal-1 loses the lectin activity, however, promotes axonal regeneration and cell transformation. We propose that As+3 binds gal-1 through sulfhydryl groups and results in oxidation of gal-1; thereafter, this oxidized gal-1 exerts its biological effect in CHOA cells. Our current results also suggest that gal-1-transfected mammalian cells uptake and accumulate more As3+. The gal-1 and gal-As3+ complexes were prepared from the cell extracts of gal-1-transfected E. coli. The circular dichroism (CD) spectrum of gal-As3+ exhibited a negative signal around 200-205 nm, whereas that of the gal-1 showed a negative signal around 205-210 nm. This shift in the CD spectrum is indicative of a substantial change in secondary structure resulted from the binding of As3+ to gal-1 proteins. Furthermore, the Isothermal titration microcalometric (ITC)studies showed that the absorption enthalpy (△Habs) of As3+ titration into gal-1 protein solution is around –55~-69 kJ/mole As3+. These adsorption enthalpy is high enough for As3+ and gal-1 to form a coordination compounds (~21 kJ/mole). The current results also showed that the gal-As3+ complexes lose partially the lectin activity and effect the cell survival of HFW cells.

李麗雯. 2001. Galectin-1蛋白與亞砷酸鈉毒性與結合作用之研究. 國立中央大學碩士論文.
林健成. 2001. 蛋白質與疏水性基材間交互作用的溫度效應及其吸附機制探討. 國立中央大學碩士論文.
陳怡均. 2001. Galectin-1與Thioredoxin peroxidase II基因之選殖及表現. 國立中央大學碩士論文.
Allen, H.J., D. Sucato, B. Woynarowska, S. Gottstine, A. Sharma, and R.J. Bernacki. 1990. Role of galaptin in ovarian carcinoma adhesion to extracellular matrix in vitro. J. Cell Biolchem. 43:43-57.
Barondes, S.H., V. Castronovo, D.N. Cooper, R.D. Cummings, K. Drickamer, T. Feizi, M.A. Gitt, J. Hirabayashi, C. Hughes, K. Kasai, and et al. 1994a. Galectins: a family of animal beta-galactoside-binding lectins. Cell. 76:597-8.
Barondes, S.H., D.N. Cooper, M.A. Gitt, and H. Leffler. 1994b. Galectins. Structure and function of a large family of animal lectins. J Biol Chem. 269:20807-10.
Bates, M.N., A.H. Smith, and C. Hopenhayn-Rich. 1992. Arsenic ingestion and internal cancers: A review. Am. J. Epidemiol. 135:462-476.
Berolero, F., G. Pozzi, E. Sabbioni, and U. Saffiotti. 1987. Cellular uptake and metabolic reduction of pentavalent to trivalent arsenic as determinants of cytotoxicity and morphological transformation. Carcinogenesis. 8:803-8.
Bianchet, M.A., H. Ahmed, G.R. Vasta, and L.M. Amzel. 2000. Soluble beta-galactosyl-binding lectin (galectin) from toad ovary: crystallographic studies of two protein-sugar complexes. Proteins. 40:378-88.
Buchet, J.P., and R. Lauwerys. 1985. Study of inorganic arsenic methylation by rat liver in vitro: relevance for the interpretation of observation in man. Arch. Toxicol.57:125-9.
Chen, C.J., C.W. Chen, M.M. Wu, and T.L. Kuo. 1992. Cancer potential in liver, lung, bladder and kidney due to ingested inorganic arsenic in drinking water. Br. J. Cancer. 66:888-892.
Chen, G.Q., J. Zhu, X.G. Shi, J.H. Ni, H.J. Zhong, G.Y. Si, X.L. Jin, W. Tang, X.S. Li, S.M. xong, Z.X. Shen, G.L. Sun, J. Ma, P. Zhang, T.D. Zhang, C. Gazin, T. Naoe, S.J. Chen, Z.Y. Wang, and Z. Chen. 1996. In virto studies on cellular and molecular mechanisms of arsenic trioxide(As2O3) in the treatment of acute promyelocytic leukemia:As2O3 induces NB4 cell apoptosis with downregulation of Bcl-2 expression and modulation of PML-RAR alpha/PML proteins. Blood. 88:1052-61.
Chen, M.L., R.S. King, and H.J. Armbrecht. 1990. Sodium-dependent phosphate transport in primary cultures of renal tubule cells from young and adult rats. J. Cell. Physiol.143:488-93.
Chiou, H.Y., Y.M. Hsueh, K.F. Liaw, S.F. Horng, M.H. Chiang, Y.S. Pu, J.S.N. Lin, C.H. Huang, and C.J. Chen. 1995. Incidence of internal cancers and ingested inorganic arsenic: A seven-year follow-up study in Taiwan. Cancer Res.55:1296-1300.
Cho, M., and R.D. Cummings. 1995a. Galectin-1, a beta-galactoside-binding lectin in Chinese hamster ovary cells. I. Physical and chemical characterization. J Biol Chem. 270:5198-206.
Cho, M., and R.D. Cummings. 1995b. Galectin-1, a beta-galactoside-binding lectin in Chinese hamster ovary cells. II. Localization and biosynthesis. J Biol Chem. 270:5207-12.
Choi, J.Y., A.J. Van Wijnen, F. Asiam, J.D. Leszyk, J.L. Stein, G.S. Stein, J.B. Lian, and S. Penman. 1998. Developmental association of the beta-galcetoside binding protein galectin-1 with the nuclear matrix of rat calvarial osteoblasts. J. Cell Sci.111:3035-3043.
Cindolo, L., G. Benvenuto, P. Salvatore, R. Pero, G. Salvatore, V. Mirone, D. Prezioso, V. Altieri, C.B. Bruni, and L. Chiariotti. 1999. Galectin-1 and galectin-3 expression in human bladder transitional-cell carcinomas. Int. J. Cancer. 84:39-43.
Cooper, D.N.W., and S.H. Barondes. 1990. Evidence for export of a muscle lectin from cytosol to extracellular matrix and for anovel secretory. J. Cell Biol.110:1681-1691.
Cooper, D.N.W., S.M. Massa, and S.H. Barondes. 1991. Endogenous muscle lectin inhibits myoblast adhesion to laminin. J. Cell Biol.115:1437-1448.
Delnomdedieu, M., M.M. Basti, J.D. Otvos, and D.J. Thomas. 1994. Reduction and binding of arsenate and dimethylarsinate by glutathione: a magnetic resonance study. Chem. Biol. Interact.90:139-55.
Dodd, J., and T.M. Jesell. 1986. Cell surface glycoconjugates and carbohydrate-binding proteins: possible recognition signals in sensory neuron development. J. Exp. Biol.124:225-238.
Fan, S.R., I.C. Ho, F.L. Yeoh, C.J. Lin, and T.C. Lee. 1996. Squalene inhibits sodium arsenite-induced sister chromatid exchanges and micronuclei in Chinese hamster ovary-K1 cells. Mutat. Res. 368:165-9.
Fischer, A.B., J.P. Buchet, and R. Lauwerys. 1985. Arsenic uptake, cytotoxicity and detoxification studied in mammalian cells in culture. Arch. Toxicol.57:168-72.
Gorbsky, G.J. 1994. Arsenic in human medicine, arsenic in the environment. Part2: Human health and ecosystem effects, New York.
Horie, H., and T. Kadoya. 2000. Identification of oxidized galectin-1 as an initial repair regulatory factor after axotomy in peripheral nerves. Neurosci Res. 38:131-7.
Horie, H., T. Kadoya, Y. Inagaki, and Y. Sohma. 2000. [Oxidized galectin-1 is a new type factor to promote nerve regeneration]. Seikagaku. 72:1245-9.
Huang, R.N., and T.C. Lee. 1996. Cellular uptake of trivalent arsenite and pentavalent arsenate in KB cells cultured in phosphate-free medium. Toxicol. Appl. Pharmacol.136:243-9.
Hughtes, R.C. 1999. Secretion of the galectin family of mammalian carbonhydrate-binding protein. Biochim. Biophys. Acta. 1473:172-185.
Hynes, M.A., L.B. Buck, M. Gitt, S. Barondes, J. Dodd, and T.M. Jesell. 1989. Carbonhydrate recognition in neuronal development: structure, expression of surface oligosaccarides and beta-galactoside-binding lectins. Clin. Symp.145:189-210.
Iglesias, M.M., G.A. Rabinovich, V. Ivanovic, C. Sotomayor, and C. Wolfenstein-Todel. 1998. Galectin-1 ovine placenta amino-acid sequence, physicochemical properties and implications in T-cell death. Eur. J. Biochem. 252:400-7.
Inagaki, Y., Y. Sohma, H. Horie, R. Nozawa, and T. Kadoya. 2000. Oxidized galectin-1 promotes axonal regeneration in peripheral nerves but does not possess lectin properties. Eur. J. Biochem. 267:2955-64.
Joubert, R., M. Caron, and D. Bladier. 1987. Brain lectin-mediated agglutinability of dissociated cells from embryonic and postnatal mouse brain. Brain Res. 433:146-150.
Kapahi, P., T. Takahashi, G. Natoli, S.R. Adams, Y. Chen, R.Y. Tsien, and M. Karin. 2000. Inhibition of NF-kB activation by arsenite through reaction with a critical cysteine in the activation loop of IkB kinase. J. Biol. Chem. 275:36062-36066.
Lee, T.C., and I.C. Ho. 1995. Modulation of cellular antioxidant defense activities by sodium arsenite in human fibroblasts. Arch. Toxicol. 69:498-504.
Li, W., and I.N. Chou. 1992. Effects of sodium arsenite on the cytoskeleton and cellular glutathione levels in cultured cells. Toxicol. Appl. Pharmacol.114:132-9.
Liu, Y.C., and H. Huang. 1997. Involvement of calcium-dependent protein kinase C in arsenite-induced genotoxicity in Chinese hamster ovary cells. J. Cell Biochem.64:423-33.
Mathews, C.K., and K.E. van Holde. 1996. Biochemistry.The Benjamin/Cummings Publishing company, Inc. Menlo Park, CA.
Mazurek, N., J. Conklin, J.C. Byrd, A. Raz, and R.S. Bresalier. 2000. Phosphorylation of the beta-galactoside-binding protein galectin-3 modulates binding to its ligands. J. Biol. Chem. 275:36311-15.
Menzel, D.B., H.K. Hamadeh, E. Lee, D.M. Meacher, V. Said, R.E. Rasmussen, H. Greene, and R.N. Roth. 1999. Arsenic binding proteins from human lymophoblastoid cells. Toxicol. Lett.105:89-101.
Moiseeva, E.P., Q. Javed, E.L. Spring, and D.P. de Bono. 2000. Galectin 1 is involved in vascular smooth muscle cell proliferation. Cardiovasc Res. 45:493-502.
Perillo, N.L., M.E. Marcus, and L.G. Baum. 1998. Galectins: versatile modulators of cell adhesion, cell proliferation, and cell death. J. Mol. Med. 76:402-12.
Perillo, N.L., K.E. Pace, J.J. Seihamer, and L.G. Baum. 1995. Apoptosis of T cells mediated by galectin-1. Nature. 78:736-739.
Tseng, W.P. 1977. Effects and dose-response relationships of skin cancer and blackfoot disease eith arsenic. Environ. Health Perspect. 19:109-119.
Vahter, M., and J. Envall. 1983. In vivo reduction of arsenate in mice and rabbits. Environ. Res.32:14-24.
Vogt, B.L., and T.G. Rossman. 2001. Effects of arsenite on p53, p21 and cyclin D expression in normal human fibroblasts --- a possible mechanism for arsenite''s comutagenicity. Mutat. Res. 478:159-68.
Wang, H.F., and T.C. Lee. 1993. Glutathione s-transferase facilitates the excretion of arsenic from arsenic-resistant chinese hamster ovary cells. Biochem. Biophys. Res. Commun.192:1093-99.
Wang, T.S., C.F. Kuo, K.Y. Jan, and H. Huang. 1996. Arsenite induces apoptosis in Chinese hamster ovary cells by generation of reactive oxygen species. J. Cell. Physiol.169:256-68.
Wen, W.N., T.L. Lieu, H.J. Chang, S.W. Wuu, M.L. Yau, and K.Y. Jan. 1981. Baseline and sodium arsenite-induced sister chromatid exchange in cultured lympocytes from patients with Blackfoot disease and healthy persons. Hum. Genet.59:201-3.
Yamanaka, K., K. Mishima, Y. Nagashima, A. Asai, Y. Sanai, and T. Kirino. 2000. Expression of galectin-1 mRNA correlates with the malignant potential of Human Gliomas and expression of antisense galectin-1 inhibits the growth of 9 Glioma cells. J. Neurosci. Res.59:722-730.
Yamaoka, K., A. Ingendoh, S. Tsubuki, Y. Nagai, and Y. Sanai. 1996. Structural and functional characterization of a novel tumor-derived rat galectin-1 having transforming growth factor (TGF) activity: the relationship between intramolecular disulfide bridges and TGF activity. J. Biochem. Tokyo. 119:878-86.
Yamauchi, H., K. Takahashi, M. Mashiko, and Y. Yamamura. 1989. Biological monitoring of arsenic exposure of gallium arsenide- and inorganic arsenic-exposed workers by determination of inorganic arsenic and its metabolites in urine and hair. Am. Ind. Hyg. Assoc. J. 50:606-12.
Yih, L.H., and T.C. Lee. 1999. Effects of exposure protocols on induction of kinetochore-plus and -minus micronuclei by arsenite in diploid human fibroblasts. Mutat. Res. 440:75-82.