Purification and partial structural and kinetic characterization of tyrosine aminotransferase from epimastigotes of Trypanosoma cruzi - PubMed (original) (raw)
Comparative Study
. 1993 Jun 15;292 ( Pt 3)(Pt 3):901-6.
doi: 10.1042/bj2920901.
Affiliations
- PMID: 8100416
- PMCID: PMC1134199
- DOI: 10.1042/bj2920901
Comparative Study
Purification and partial structural and kinetic characterization of tyrosine aminotransferase from epimastigotes of Trypanosoma cruzi
M Montemartini et al. Biochem J. 1993.
Abstract
Tyrosine aminotransferase was purified to homogeneity from epimastigotes of Trypanosoma cruzi by a method involving chromatography on DEAE-cellulose, gel filtration on Sephacryl S-200 and chromatography on Mono Q in an f.p.l.c. system. The purified enzyme showed a single band in SDS/PAGE, with an apparent molecular mass of 45 kDa. Since the apparent molecular mass of the native enzyme, determined by gel filtration, is 91 kDa, the native enzyme is a dimer of similar subunits. The amino-acid composition was determined, as well as the sequences of three internal peptides obtained by CNBr cleavage at Met residues. Both criteria suggest considerable similarity with the tyrosine aminotransferases from rat and from human liver. The enzyme contains nine 1/2 Cys residues, three free and the others forming three disulphide bridges. The enzyme is not N-glycosylated. The isoelectric point is 4.6-4.8. The optimal pH for the reaction of the enzyme with tyrosine as a substrate is 7.0. The apparent Km values for tyrosine, phenylalanine and tryptophan, with pyruvate as a co-substrate, were 6.8, 17.9 and 21.4 mM, respectively, whereas those for pyruvate, alpha-oxoglutarate and oxaloacetate, with tyrosine as a substrate, were 0.5, 38 and 16 mM respectively. The purified tyrosine aminotransferase acts as an alanine aminotransferase as well and the activity seems to reside in the same enzyme molecule. The results suggest that the enzyme is a general aromatic-amino-acid transaminase, with high sequence similarity to tyrosine aminotransferases from rat and human liver.
Similar articles
- Purification and partial structural and kinetic characterization of an aromatic L-alpha-hydroxy acid dehydrogenase from epimastigotes of Trypanosoma cruzi.
Montemartini M, Santomé JA, Cazzulo JJ, Nowicki C. Montemartini M, et al. Mol Biochem Parasitol. 1994 Nov;68(1):15-23. doi: 10.1016/0166-6851(94)00145-6. Mol Biochem Parasitol. 1994. PMID: 7891739 - Isolation and partial characterization of a broad specificity aminotransferase from Leishmania mexicana promastigotes.
Vernal J, Cazzulo JJ, Nowicki C. Vernal J, et al. Mol Biochem Parasitol. 1998 Oct 30;96(1-2):83-92. doi: 10.1016/s0166-6851(98)00117-0. Mol Biochem Parasitol. 1998. PMID: 9851609 - A recombinant tyrosine aminotransferase from Trypanosoma cruzi has both tyrosine aminotransferase and alanine aminotransferase activities.
Montemartini M, Búa J, Bontempi E, Zelada C, Ruiz AM, Santomé JA, Cazzulo JJ, Nowicki C. Montemartini M, et al. FEMS Microbiol Lett. 1995 Nov 1;133(1-2):17-20. doi: 10.1111/j.1574-6968.1995.tb07854.x. FEMS Microbiol Lett. 1995. PMID: 8566704 - Presence and subcellular localization of tyrosine aminotransferase and p-hydroxyphenyllactate dehydrogenase in epimastigotes of Trypanosoma cruzi.
Nowicki C, Montemartini M, Duschak V, Santomé JA, Cazzulo JJ. Nowicki C, et al. FEMS Microbiol Lett. 1992 Apr 15;71(2):119-24. doi: 10.1016/0378-1097(92)90498-d. FEMS Microbiol Lett. 1992. PMID: 1351016 - Recombinant tyrosine aminotransferase from Trypanosoma cruzi: structural characterization and site directed mutagenesis of a broad substrate specificity enzyme.
Nowicki C, Hunter GR, Montemartini-Kalisz M, Blankenfeldt W, Hecht H, Kalisz HM. Nowicki C, et al. Biochim Biophys Acta. 2001 Apr 7;1546(2):268-81. doi: 10.1016/s0167-4838(01)00136-4. Biochim Biophys Acta. 2001. PMID: 11295433
Cited by
- Uptake of l-Alanine and Its Distinct Roles in the Bioenergetics of Trypanosoma cruzi.
Girard RMBM, Crispim M, Alencar MB, Silber AM. Girard RMBM, et al. mSphere. 2018 Jul 18;3(4):e00338-18. doi: 10.1128/mSphereDirect.00338-18. mSphere. 2018. PMID: 30021880 Free PMC article. - Role of Δ1-pyrroline-5-carboxylate dehydrogenase supports mitochondrial metabolism and host-cell invasion of Trypanosoma cruzi.
Mantilla BS, Paes LS, Pral EM, Martil DE, Thiemann OH, Fernández-Silva P, Bastos EL, Silber AM. Mantilla BS, et al. J Biol Chem. 2015 Mar 20;290(12):7767-90. doi: 10.1074/jbc.M114.574525. Epub 2015 Jan 26. J Biol Chem. 2015. PMID: 25623067 Free PMC article. - Structure of tyrosine aminotransferase from Leishmania infantum.
Moreno MA, Abramov A, Abendroth J, Alonso A, Zhang S, Alcolea PJ, Edwards T, Lorimer D, Myler PJ, Larraga V. Moreno MA, et al. Acta Crystallogr F Struct Biol Commun. 2014 May;70(Pt 5):583-7. doi: 10.1107/S2053230X14007845. Epub 2014 Apr 25. Acta Crystallogr F Struct Biol Commun. 2014. PMID: 24817714 Free PMC article. - Branched-chain and aromatic amino acid catabolism into aroma volatiles in Cucumis melo L. fruit.
Gonda I, Bar E, Portnoy V, Lev S, Burger J, Schaffer AA, Tadmor Y, Gepstein S, Giovannoni JJ, Katzir N, Lewinsohn E. Gonda I, et al. J Exp Bot. 2010 Feb;61(4):1111-23. doi: 10.1093/jxb/erp390. Epub 2010 Jan 11. J Exp Bot. 2010. PMID: 20065117 Free PMC article. - Database of Trypanosoma cruzi repeated genes: 20,000 additional gene variants.
Arner E, Kindlund E, Nilsson D, Farzana F, Ferella M, Tammi MT, Andersson B. Arner E, et al. BMC Genomics. 2007 Oct 26;8:391. doi: 10.1186/1471-2164-8-391. BMC Genomics. 2007. PMID: 17963481 Free PMC article.
References
- Biochemistry. 1967 Jul;6(7):1948-54 - PubMed
- Nature. 1957 Dec 21;180(4599):1425 - PubMed
- Biochem Biophys Res Commun. 1969 May 22;35(4):467-73 - PubMed
- Nature. 1970 Aug 15;227(5259):680-5 - PubMed
- Biochim Biophys Acta. 1971 Jan 26;230(1):137-45 - PubMed
Publication types
MeSH terms
Substances
LinkOut - more resources
Full Text Sources
Molecular Biology Databases
Miscellaneous