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.
Skip Nav Destination
Follow us on Twitter @Biochem_Journal
Article navigation
March 1998
-
Cover Image
Cover Image
- PDF Icon PDF LinkFront Matter
- PDF Icon PDF LinkTable of Contents
Research Article|
March 01 1998
The NADP+-linked glutamate dehydrogenase from Trypanosoma cruzi: sequence, genomic organization and expression Available to Purchase
Patricia BARDERI;
Patricia BARDERI
*Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín. Av. General Paz y Albarellos, Casilla de Correo 30, 1650 San Martín, Prov. de 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
Search for other works by this author on:
Oscar CAMPETELLA;
Oscar CAMPETELLA
*Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín. Av. General Paz y Albarellos, Casilla de Correo 30, 1650 San Martín, Prov. de 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
Search for other works by this author on:
Alberto Carlos C. FRASCH;
Alberto Carlos C. FRASCH
*Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín. Av. General Paz y Albarellos, Casilla de Correo 30, 1650 San Martín, Prov. de 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
Search for other works by this author on:
José A. SANTOMÉ;
José A. SANTOMÉ
‡IQUIFIB (UBA-CONICET), Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junin 956, 1113 Buenos Aires, Argentina
Search for other works by this author on:
Ulf HELLMAN;
Ulf HELLMAN
§Ludwig Institute for Cancer Research, Uppsala Branch, Box 595, S-751 24 Uppsala, Sweden
Search for other works by this author on:
Ulf PETTERSSON;
Ulf PETTERSSON
ǁDepartment of Medical Genetics, Biomedical Center, Uppsala University, Box 589, S-751 24 Uppsala, Sweden
Search for other works by this author on:
Juan José CAZZULO
Juan José CAZZULO
1
*Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín. Av. General Paz y Albarellos, Casilla de Correo 30, 1650 San Martín, Prov. de 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
1To whom correspondence should be addressed at: Instituto de Investigaciones Biotecnológicas, Universidad Nacional de General San Martín. Av. General Paz y Albarellos, Casilla de Correo 30, 1650 San Martín, Prov. de Buenos Aires, Argentina.
Search for other works by this author on:
Publisher: Portland Press Ltd
Received:
July 10 1997
Revision Received:
November 10 1997
Accepted:
November 19 1997
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1998
1998
Biochem J (1998) 330 (2): 951–958.
Article history
Received:
July 10 1997
Revision Received:
November 10 1997
Accepted:
November 19 1997
Citation
Patricia BARDERI, Oscar CAMPETELLA, Alberto Carlos C. FRASCH, José A. SANTOMÉ, Ulf HELLMAN, Ulf PETTERSSON, Juan José CAZZULO; The NADP+-linked glutamate dehydrogenase from Trypanosoma cruzi: sequence, genomic organization and expression. Biochem J 1 March 1998; 330 (2): 951–958. doi: https://doi.org/10.1042/bj3300951
Download citation file:
Sign in
Don't already have an account? Register
Sign in to your personal account
You could not be signed in. Please check your email address / username and password and try again.
Could not validate captcha. Please try again.
Biochemical Society Member Sign in
Sign InSign in via your Institution
Sign in via your InstitutionGet Access To This Article
Follow us on Twitter @Biochem_Journal
Open Access for all
We offer compliant routes for all authors from 2025. With library support, there will be no author nor reader charges in 5 journals. Check here |
![]() View past webinars > |