Heparan sulfate, heparin, and heparinase activity detection on polyacrylamide gel electrophoresis using the fluorochrome tris(2,2′-bipyridine) ruthenium

Mostrar otras versiones (1)

The paper shows the ability of the fluorochrome tris(2,2′-bipyridine) ruthenium (II) (Rubipy) to detect heparan sulfate, heparin, and heparinase activity of M3 murine mammary adenocarcinoma cells as well as bacterial heparinases I, II, and III in native polyacrylamide gel electrophoresis (PAGE). The...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autor principal: Rozenberg, G.I
Otros Autores: Espada, J., de Cidre, L.L, Eiján, A.M, Calvo, J.C, Bertolesi, G.E
Formato: Capítulo de libro
Lenguaje:Inglés
Publicado: 2001
Acceso en línea:Registro en Scopus
DOI
Handle
Registro en la Biblioteca Digital
Aporte de:Registro referencial: Solicitar el recurso aquí
Biblioteca Central Dr. Luis F. Leloir (FCEN) de Universidad de Buenos Aires
LEADER 09440caa a22012737a 4500
001 PAPER-20683
003 AR-BaUEN
005 20230518205206.0
008 190411s2001 xx ||||fo|||| 00| 0 eng|d
024 7 |2 scopus  |a 2-s2.0-0035165166 
024 7 |2 cas  |a 2,2' bipyridine, 366-18-7; fluorochrome, 37332-03-9; heparan sulfate, 9050-30-0; heparin, 37187-54-5, 8057-48-5, 8065-01-8, 9005-48-5; heparin lyase, 37290-85-0, 9025-39-2; tris(2,2' bipyridine)ruthenium, 14323-06-9; 2,2'-Dipyridyl, 366-18-7; Fluorescent Dyes; Heparin, 9005-49-6; Heparin Lyase, 4.2.2.7; Heparitin Sulfate, 9050-30-0; Polysaccharide-Lyases, 4.2.2.-; heparinase II, 4.2.2.-; heparitinsulfate lyase, 4.2.2.8; tris(2,2'-bipyridine)ruthenium II, 15158-62-0 
040 |a Scopus  |b spa  |c AR-BaUEN  |d AR-BaUEN 
030 |a ELCTD 
100 1 |a Rozenberg, G.I. 
245 1 0 |a Heparan sulfate, heparin, and heparinase activity detection on polyacrylamide gel electrophoresis using the fluorochrome tris(2,2′-bipyridine) ruthenium 
260 |c 2001 
270 1 0 |m Rozenberg, G.I.; Depto. de Química Biol., Fac. de Ciencias Exact. y Naturales, Universidad de Buenos Aires, Buenos Aires, Argentina; email: garozenb@qb.fcen.uba.ar 
506 |2 openaire  |e Política editorial 
504 |a Casu, B., (1985) Adv. Carbohydr. Chem. Biochem., 43, pp. 51-134 
504 |a Linhardt, R.J., Turnbull, J.E., Wang, H.M., Loganathan, D., Gallagher, J.T., (1990) Biochem., 29, pp. 2611-2617 
504 |a Jandik, K.A., Gu, K., Linhardt, R.J., (1994) Glycobiol., 4, pp. 289-296 
504 |a Matzner, Y., Bar-Ner, M., Yahalom, J., Ishai-Michaeli, R., Fuks, Z., Voldavsky, I., (1985) J. Clin. Invest., 76, pp. 1306-1313 
504 |a Bashkin, P., Razin, E., Eldor, A., Vlodavsky, I., (1990) Blood, 75, pp. 2204-2212 
504 |a Vlodavsky, I., Eldor, A., Hairnovitz-Friedman, A., Matzner, Y., Ishai-Michaeli, R., Lider, O., Naparstek, Y., Fuks, Z., (1992) J. Invas. Metast., 12, pp. 112-127 
504 |a Nakajima, M., Irimura, T., Nicolson, G.L., (1988) J. Cell. Biochem., 36, pp. 157-167 
504 |a Jin, L., Nakajima, M., Nicolson, G.L., (1990) Int. J. Cancer, 45, pp. 1088-1095 
504 |a Vlodavsky, I., Friedmann, Y., Elkin, M., Aingorn, H., Atzmon, R., Ishai-Michaeli, R., Bian, M., Pecker, I., (1999) Nature Med., 5, pp. 793-802 
504 |a Hulett, M.D., Freeman, C., Hamdorf, B.J., Baker, R.T., Harris, M.J., Parish, C.R., (1999) Nature Med., 5, pp. 803-809 
504 |a Nakajima, M., Irimura, T., Di Ferrante, D., Di Ferrante, N., Nicolson, G.L., (1983) Science, 220, pp. 611-613 
504 |a De Vouge, M.W., Yamazaki, A., Bennett, S.A., Chen, J.H., Shwed, P.S., Couture, C., Birnboim, H.C., (1994) Int. J. Cancer, 56, pp. 286-294 
504 |a Al-Hakim, A., Linhardt, R.J., (1991) Appl. Theor. Electrophor., 1, pp. 305-312 
504 |a Linker, A., Hovingh, P., (1972) Biochemistry, 11, pp. 563-568 
504 |a Bertolesi, G.E., Trigoso, C.I., Espada, J., Stockert, J.C., (1995) J. Histochem. Cytochem., 43, pp. 537-543 
504 |a Bertolesi, G.E., Stockert, J.C., Lauría de Cidre, L., (1997) Int. J. Oncol., 11, pp. 1221-1225 
504 |a Kelly, J.M., Tossi, A.M., McConnell, D.J., OhUigin, C., (1985) Nucleic Acids Res., 13, pp. 6017-6034 
504 |a Bitter, T., Muir, H.M., (1962) Anal. Biochem., 4, pp. 330-334 
504 |a Laemmli, U.K., (1970) Nature, 227, pp. 680-685 
504 |a Munson, P.J., Rodbard, D., (1980) Anal. Biochem., 107, pp. 220-239 
504 |a Bal de Kier Joffé, E., Puricelli, L., Vidal, M.C., Sacerdote de Lustig, E., (1983) J. Exp. Clin. Cancer Res., 2, pp. 151-160 
504 |a Bal de Kier Joffé, E., Puricelli, L., Sacerdote de Lustig, E., (1986) Invasion Metastasis, 6, pp. 302-312 
504 |a Johnson, E.A., Mulloy, B., (1976) Carbohydr. Res., 51, pp. 119-127 
504 |a Tas, J., Roozemond, R.C., (1973) Histochem. J., 5, pp. 425-436 
504 |a Stone, A.L., Bradley, D.F., (1967) Biochim Biophys. Acta, 148, pp. 172-192 
504 |a Desai, U.R., Wang, H., Ampofo, S.A., Linhardt, R.J., (1993) Anal. Biochem., 213, pp. 120-127 
504 |a Rusnati, M., Tulipano, G., Spillman, D., Tanghetti, E., Oreste, P., Zoppetti, G., Gliacca, M., Presta, M.J., (1999) Biol. Chem., 274, pp. 28198-28205 
504 |a Nader, H.B., Porcionatto, M.A., Tersariol, I.L.S., Pinhal, M.A.S., Oliveira, F.W., Moraes, C.T., Dietrich, C.P., (1990) J. Biol. Chem., 265, pp. 16807-16813 
504 |a Sasisekharan, R., Bulmer, M., Moremen, K.W., Cooney, C.L., Langer, R., (1993) Proc. Natl. Acad. Sci. USA, 90, pp. 3660-3664 
504 |a Bar-Ner, M., Eldor, A., Wasserman, L., Matzner, Y., Cohen, I.R., Fuks, Z., Vlodavsky, I., (1987) Blood, 70, pp. 551-557 
504 |a Bertolesi, G.E., Lauría de Cidre, L., Sacerdote de Lustig, E., Eiján, A.M., (1995) Cancer Lett., 90, pp. 123-131 
504 |a Brzu, T., Pascal, M., Maman, M., Roque, C., Lafont, F., Rousselet, A., (1996) J. Cell. Physiol., 167, pp. 8-21 
504 |a Khan, M.Y., Newman, S.A., (1991) Anal. Biochem., 196, pp. 373-376 
520 3 |a The paper shows the ability of the fluorochrome tris(2,2′-bipyridine) ruthenium (II) (Rubipy) to detect heparan sulfate, heparin, and heparinase activity of M3 murine mammary adenocarcinoma cells as well as bacterial heparinases I, II, and III in native polyacrylamide gel electrophoresis (PAGE). The technique is based on the electrophoretic mobility of high molecular weight heparins and subsequent staining with Rubipy (50 μg/mL). The minimum content of heparin detected by fluorescence in a UV transilluminator was 25-50 ng. The number of Rubipy molecules bound to heparin, determined in relationship to the number of disaccharide units (DU), showed that two to six heparin disaccharide units are bound by each fluorochrome molecule. Scatchard plot analysis showed one Rubipy-binding site (Kd= (8.56 ± 2.97) × 10-5 M). Heparinase activity was determined by densitometric analysis of the fluorescence intensity of the heparin-containing band of the gel. While heparinase I (EC 4.2.2.7.) degraded heparin and, to a lower degree, partially N-desulfated N-acetylated heparin (N-des N-Ac), heparinase II (no EC number) could efficiently degrade heparan sulfate (HS) and partially N-des N-Ac heparin. Finally, heparinase III (EC 4.2.2.8.) degraded HS almost exclusively. Only heparin and N-des N-Ac heparin were substrates for M3 tumor cell heparinases. We describe a qualitative, sensitive and simple method to detect heparinase activity and determine its substrate specificity using Rubipy fluorescence with heparin and heparan sulfate in multiple biological samples tested in parallel.  |l eng 
593 |a Departamento de Química Biológica, Argentina 
593 |a Departamento de Ciencas Biológicas, Universidad de Buenos Aires, Argentina 
593 |a Instituto de Investigaciones Biomédicas, Consejo Superior de Investigaciones Científicas, Universidad Autónoma de Madrid, Madrid, Spain 
593 |a Instituto de Oncología 'A.H.Roffo', Universidad de Buenos Aires, Argentina 
593 |a Departamento de Química Biológica, Universidad de Buenos Aires, Argentina 
690 1 0 |a HEPARAN SUTFATE 
690 1 0 |a HEPARIN 
690 1 0 |a HEPARINASE 
690 1 0 |a TRIS (2,2′BIPYRIDINE) RUTHENIUM (II) EL 4139 
690 1 0 |a 2,2' BIPYRIDINE 
690 1 0 |a FLUOROCHROME 
690 1 0 |a HEPARAN SULFATE 
690 1 0 |a HEPARIN 
690 1 0 |a HEPARIN LYASE 
690 1 0 |a RUTHENIUM COMPLEX 
690 1 0 |a TRIS(2,2' BIPYRIDINE)RUTHENIUM 
690 1 0 |a UNCLASSIFIED DRUG 
690 1 0 |a ADENOCARCINOMA 
690 1 0 |a ANALYTIC METHOD 
690 1 0 |a ANIMAL CELL 
690 1 0 |a ARTICLE 
690 1 0 |a BINDING SITE 
690 1 0 |a CARCINOMA CELL 
690 1 0 |a DENSITOMETRY 
690 1 0 |a ELECTROPHORETIC MOBILITY 
690 1 0 |a ENZYME ACTIVITY 
690 1 0 |a ENZYME SPECIFICITY 
690 1 0 |a FLUORESCENCE 
690 1 0 |a NONHUMAN 
690 1 0 |a POLYACRYLAMIDE GEL ELECTROPHORESIS 
690 1 0 |a 2,2'-DIPYRIDYL 
690 1 0 |a ADENOCARCINOMA 
690 1 0 |a ANIMALS 
690 1 0 |a ELECTROPHORESIS, POLYACRYLAMIDE GEL 
690 1 0 |a FEMALE 
690 1 0 |a FLUORESCENT DYES 
690 1 0 |a HEPARIN 
690 1 0 |a HEPARIN LYASE 
690 1 0 |a HEPARITIN SULFATE 
690 1 0 |a MAMMARY NEOPLASMS, ANIMAL 
690 1 0 |a MICE 
690 1 0 |a MOLECULAR STRUCTURE 
690 1 0 |a POLYSACCHARIDE-LYASES 
690 1 0 |a TUMOR CELLS, CULTURED 
700 1 |a Espada, J. 
700 1 |a de Cidre, L.L. 
700 1 |a Eiján, A.M. 
700 1 |a Calvo, J.C. 
700 1 |a Bertolesi, G.E. 
773 0 |d 2001  |g v. 22  |h pp. 3-11  |k n. 1  |p Electrophoresis  |x 01730835  |w (AR-BaUEN)CENRE-4560  |t Electrophoresis 
856 4 1 |u https://www.scopus.com/inward/record.uri?eid=2-s2.0-0035165166&doi=10.1002%2f1522-2683%28200101%2922%3a1%3c3%3a%3aAID-ELPS3%3e3.0.CO%3b2-G&partnerID=40&md5=7d42eba62024137e9c5d0969f46510f0  |y Registro en Scopus 
856 4 0 |u https://doi.org/10.1002/1522-2683(200101)22:1<3::AID-ELPS3>3.0.CO;2-G  |y DOI 
856 4 0 |u https://hdl.handle.net/20.500.12110/paper_01730835_v22_n1_p3_Rozenberg  |y Handle 
856 4 0 |u https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_01730835_v22_n1_p3_Rozenberg  |y Registro en la Biblioteca Digital 
961 |a paper_01730835_v22_n1_p3_Rozenberg  |b paper  |c PE 
962 |a info:eu-repo/semantics/article  |a info:ar-repo/semantics/artículo  |b info:eu-repo/semantics/publishedVersion 
999 |c 81636 

Ejemplares similares