Abstract:
NADP-linked glutamate dehydrogenase (NADP+-GluDH, EC 1.4.1.4) has been purified to homogeneity from epimastigotes of Trypanosoma cruzi by an improved procedure, and the amino acid sequences of 11 internal peptides obtained by digestion with trypsin, endopeptidase Lys-C, endopeptidase Arg-C or CNBr have been obtained. Using oligonucleotide primers synthesized according to the amino acid sequence of the N-terminus of the mature enzyme and to the nucleotide sequence of a clone corresponding to the C-terminus, obtained by immunological screening of an expression library, two complete open reading frames (TcGluDH1 and TcGluDH2) were isolated and sequenced. The sequences obtained are most similar to that of the NADP+-GluDH of Escherichia coli (70-72% identity), and less similar (50-56%) to those of lower eukaryotes. Using TcGluDH1 as a probe, evidence for the presence of several genes and developmental regulation of the expression of NADP+-GluDH in different parasite stages was obtained. TcGluDH1 encodes an enzymically active protein, since its expression in E. coli resulted in the production of a GluDH activity with kinetic parameters similar to those of the natural enzyme.
Registro:
Documento: |
Artículo
|
Título: | The NADP+ -linked glutamate dehydrogenase from Trypanosoma cruzi: Sequence, genomic organization and expression |
Autor: | Barderi, P.; Campetella, O.; Frasch, A.C.C.; Santomé, J.A.; Hellman, U.; Pettersson, U.; Cazzulo, J.J. |
Filiación: | Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín, Av. Gen. Paz y Albar., C.Correo 30, Buenos Aires, Argentina Instituto de Investigaciones Bioquímicas 'Luis F. Leloir', Fundación Campomar/CONICET/Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires, A. Machado 151, 1405 Buenos Aires, Argentina IQUIFIB (UBA-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junin 956, 1113 Buenos Aires, Argentina Ludwig Institute for Cancer Research, Uppsala Branch, Box 595, S-751 24 Uppsala, Sweden Department of Medical Genetics, Biomedical Center, Uppsala University, S-751 24 Uppsala, Sweden
|
Palabras clave: | glutamate dehydrogenase; nicotinamide adenine dinucleotide phosphate; oligonucleotide; proteinase; trypsin; amino acid sequence; article; carboxy terminal sequence; epimastigote; genome; nucleotide sequence; open reading frame; priority journal; protein expression; trypanosoma cruzi; Amino Acid Sequence; Animals; Base Sequence; Cytosol; DNA, Protozoan; Escherichia coli; Evolution, Molecular; Glutamate Dehydrogenase (NADP+); Molecular Sequence Data; Recombinant Proteins; Trypanosoma cruzi; Escherichia coli; Eukaryota; Protozoa; Trypanosoma; Trypanosoma cruzi |
Año: | 1998
|
Volumen: | 330
|
Número: | 2
|
Página de inicio: | 951
|
Página de fin: | 958
|
DOI: |
http://dx.doi.org/10.1042/bj3300951 |
Título revista: | Biochemical Journal
|
Título revista abreviado: | Biochem. J.
|
ISSN: | 02646021
|
CODEN: | BIJOA
|
CAS: | DNA, Protozoan; Glutamate Dehydrogenase (NADP+), EC 1.4.1.4; Recombinant Proteins
|
Registro: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_02646021_v330_n2_p951_Barderi |
Referencias:
- Smith, E.L., Austen, B.M., Blumenthal, K.M., Nyc, J.F., (1975) Enzymes (3rd Edn.), 11, pp. 293-367
- Cazzulo, J.J., Juan, S.M., Segura, E.L., (1977) Comp. Biochem. Physiol., 56 B, pp. 301-303
- Cazzulo, J.J., Franke De Cazzulo, B.M., Higa, A.I., Segura, E.L., (1979) Comp. Biochem. Physiol., 64 B, pp. 129-131
- Juan, S.M., Segura, E.L., Cazzulo, J.J., (1978) Int. J. Biochem., 9, pp. 395-400
- Walter, R.D., Ebert, F., (1979) J. Protozool., 26, pp. 653-656
- Cazzulo, J.J., (1984) Comp. Biochem. Physiol., 79 B, pp. 309-320
- Urbina, J.A., Azavache, V., (1984) Mol. Biochem. Parasitol., 11, pp. 241-255
- Cazzulo, J.J., Nowicki, C., Santomé, J.A., Wernstedt, C., Hellman, U., (1988) FEMS Microbiol. Lett, 56, pp. 215-220
- Duschak, V.G., Cazzulo, J.J., (1991) FEMS Microbiol. Lett., 83, pp. 131-136
- Souto-Padrón, T., Campetella, O., Cazzulo, J.J., De Souza, W., (1990) J. Cell Sci., 96, pp. 485-490
- Carneiro, M., Caldas, R.A., (1983) Comp. Biochem. Physiol., 75 B, pp. 61-64
- Cazzulo, J.J., Franke De Cazzulo, B.M., Engel, J.C., Cannata, J.J.B., (1985) Mol. Biochem. Parasitol., 16, pp. 329-343
- Yee, J., Dennis, P.P., (1992) J. Biol. Chem., 267, pp. 7539-7544
- Andrews, N.W., Colli, W., (1982) J. Protozool., 29, pp. 264-269
- Cazzulo, J.J., Couso, R., Raimondi, A., Wernstedt, C., Hellman, U., (1989) Mol. Biochem. Parasitol., 33, pp. 33-41
- Fraser, R.D.B., Suzuki, E., (1973) Physical Principles and Techniques of Protein Chemistry, Part C, 21, pp. 301-355. , Leach S. J., ed., Academic Press, New York
- Bradford, M.M., (1976) Anal. Biochem., 72, pp. 248-254
- Laemmli, U.K., (1970) Nature (London), 227, pp. 680-685
- Martínez, J., Campetella, O., Frasch, A.C.C., Cazzulo, J.J., (1991) Infect. Immunol., 59, pp. 4275-4277
- Campetella, O., Martinez, J., Cazzulo, J.J., (1990) FEMS Microbiol. Lett., 67, pp. 145-150
- Renlund, S., Klintrot, I.-M., Nunn, M., Schrimsher, J.L., Wernstedt, C., Hellman, U., (1990) J. Chromatogr., 512, pp. 325-335
- Sambrook, J., Fritsch, E.T., Maniatis, T., (1989) Molecular Cloning: A Laboratory Manual, 2nd Edn., , Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
- Chomczynski, P., Sacchi, N., (1987) Anal. Biochem., 162, pp. 156-159
- Ibáñez, C., Affranchino, J.L., Frasch, A.C.C., (1987) Mol. Biochem. Parasitol., 25, pp. 175-184
- Henriksson, J., Åslund, L., Macina, R.A., Franke De Cazzulo, B.M., Cazzulo, J.J., Frasch, A.C.C., Pettersson, U., (1990) Mol. Biochem. Parasitol., 42, pp. 213-224
- Henriksson, J., Porcel, B., Ridåker, M., Ruiz, A., Sabaj, V., Galanti, N., Cazzulo, J.J., Pettersson, U., (1995) Mol. Biochem. Parasitol., 73, pp. 63-74
- McCarthy-Burke, C., Taylor, Z.A., Busch, G.A., (1989) Gene, 82, pp. 177-189
- Sanger, F., Niklen, S., Coulson, A.R., (1977) Proc. Natl. Acad. Sci. U.S.A., 74, pp. 5463-5467
- Studier, F.W., Rosenberg, A.H., Dunn, J.J., Dubendorff, J.W., (1990) Methods Enzymol., 185, pp. 60-89
- Campetella, O., Henriksson, J., Åslund, L., Frasch, A.C.C., Pettersson, U., Cazzulo, J.J., (1992) Mol. Biochem. Parasitol., 50, pp. 225-234
- Bontempi, E.J., Búa, J., Åslund, L., Porcel, B., Segura, E.L., Henriksson, J., Orn, A., Ruiz, A.M., (1993) Mol. Biochem. Parasitol., 59, pp. 253-262
- Valle, F., Becerril, B.L., Chen, E., Seeburg, P.H., Heyneker, H., Bolivar, F., (1984) Gene, 27, pp. 193-199
- Bansal, A., Dayton, M.A., Zalkin, H., Colman, R.F., (1989) J. Biol. Chem., 264, pp. 9827-9835
- Fleischmann, R.D., Adams, M.D., White, O., Clayton, R.A., Kirkness, A.R., Kerlavage, E.F., (1995) Science, 269, pp. 496-512
- Gillin, F.D., Reiner, D.S., McCaffery, J.M., (1996) Annu. Rev. Microbiol., 50, pp. 679-705
- Cock, J.M., Kim, K.D., Miller, P.W., Hutson, R.G., Schmidt, R.R., (1991) Plant Mol. Biol., 17, pp. 1023-1044
- Kinnaird, J.H., Fincham, J.R.S., (1983) Gene, 26, pp. 253-260
- Nagasu, T., Hall, B.D., (1985) Gene, 37, pp. 247-253
- Benachenhou, N., Baldacci, G., (1991) Mol. Gen. Genet., 230, pp. 345-352
- Benachenhou-Lahfa, N., Labedan, B., Forterre, P., (1994) Gene, 140, pp. 17-24
- Zelada, C., Montemartini, M., Cazzulo, J.J., Nowicki, C., (1996) Mol. Biochem Parasitol., 79, pp. 225-228
- Tomas, A.M., Kelly, J.M., (1996) Mol. Biochem. Parasitol., 76, pp. 91-103
- Bormann, E.R., Eikmanns, B.J., Sahm, H., (1992) Mol. Microbiol., 6, pp. 317-326
- De Zoysa, P.A., Connerton, I.F., Watson, D.C., Johnston, J.R., (1991) Curr. Genet., 20, pp. 219-224
- Hawkins, A.R., Gurr, S.J., Montague, P., Kinghorn, J.R., (1989) Mol. Gen. Genet., 218, pp. 105-111
- Schaap, P.J., Muller, Y., Baars, J.J., Op Den Camp, H.J., Sonnenberg, A.S., Van Griensven, L.J., Visser, J., (1996) Mol. Gen. Genet., 250, pp. 339-347
- Chavez, S., Reyes, J.C., Chauvat, F., Florencio, F.J., Candau, P., (1995) Plant Mol. Biol., 28, pp. 173-188
- Hein, J.J., (1990) Methods Enzymol., 183, pp. 626-645
Citas:
---------- APA ----------
Barderi, P., Campetella, O., Frasch, A.C.C., Santomé, J.A., Hellman, U., Pettersson, U. & Cazzulo, J.J.
(1998)
. The NADP+ -linked glutamate dehydrogenase from Trypanosoma cruzi: Sequence, genomic organization and expression. Biochemical Journal, 330(2), 951-958.
http://dx.doi.org/10.1042/bj3300951---------- CHICAGO ----------
Barderi, P., Campetella, O., Frasch, A.C.C., Santomé, J.A., Hellman, U., Pettersson, U., et al.
"The NADP+ -linked glutamate dehydrogenase from Trypanosoma cruzi: Sequence, genomic organization and expression"
. Biochemical Journal 330, no. 2
(1998) : 951-958.
http://dx.doi.org/10.1042/bj3300951---------- MLA ----------
Barderi, P., Campetella, O., Frasch, A.C.C., Santomé, J.A., Hellman, U., Pettersson, U., et al.
"The NADP+ -linked glutamate dehydrogenase from Trypanosoma cruzi: Sequence, genomic organization and expression"
. Biochemical Journal, vol. 330, no. 2, 1998, pp. 951-958.
http://dx.doi.org/10.1042/bj3300951---------- VANCOUVER ----------
Barderi, P., Campetella, O., Frasch, A.C.C., Santomé, J.A., Hellman, U., Pettersson, U., et al. The NADP+ -linked glutamate dehydrogenase from Trypanosoma cruzi: Sequence, genomic organization and expression. Biochem. J. 1998;330(2):951-958.
http://dx.doi.org/10.1042/bj3300951