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Abdian, P.L.; Lellouch, A.C.; Gautier, C.; Ielpi, L.; Geremia, R.A. "Identification of essential amino acids in the bacterial α-mannosyltransferase AceA" (2000) Journal of Biological Chemistry. 275(51):40568-40575
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Abstract:

The α-mannosyltransferase AceA from Acetobacter xylinum belongs to the CaZY family 4 of retaining glycosyltransferases. We have identified a series of either highly conserved or invariant residues that are found in all family 4 enzymes as well as other retaining glycosyltransferases. These residues included Glu-287 and Glu-295, which comprise an EX7E motif and have been proposed to be involved in catalysis. Alanine replacements of each conserved residue were constructed by site-directed mutagenesis. The mannosyltransferase activity of each mutant was examined by both an in vitro transferase assay using recombinant mutant AceA expressed in Escherichia coli and by an in vivo rescue assay by expressing the mutant AceA in a Xanthomonas campestris gumH- strain. We found that only mutants K211A and E287A lost all detectable activity both in vitro and in vivo, whereas E295A retained residual activity in the more sensitive in vivo assay. H127A and S162A each retained reduced but significant activities both in vitro and in vivo. Secondary structure predictions of AceA and subsequent comparison with the crystal structures of the T4 β-glucosyltransferase and MurG suggest that AceA Lys-211 and Glu-295 are involved in nucleotide sugar donor binding, leaving Glu-287 of the EX7E as a potential catalytic residue.

Registro:

Documento: Artículo
Título:Identification of essential amino acids in the bacterial α-mannosyltransferase AceA
Autor:Abdian, P.L.; Lellouch, A.C.; Gautier, C.; Ielpi, L.; Geremia, R.A.
Filiación:Instituto de Investigaciones Bioquímicas Fundación Campomar, Facultad de Ciencias Exactas y Naturales, Consejo Nacional de Investigaciones Científicas y Técnicas, Avenida Patricias Argentinas 435, 1045 Buenos Aires, Argentina
Centre de Recherches Sur Les Macromolécules Végétales, CNRS, Joseph Fourier University, BP 53X, 38041 Grenoble Cedex 9, France
CERMAV/CNRS, BP 53, 38041 Grenoble Cedex 9, France
Carrera del Investigador, Consejo Nacional de Investigaciones CientIficas y Técnicas, Buenos Aires, Argentina
Palabras clave:alpha mannosyltransferase acea; bacterial enzyme; essential amino acid; glucosyltransferase; mannosyltransferase; unclassified drug; Acetobacter; amino acid substitution; article; cellular distribution; enzyme activity; enzyme analysis; enzyme structure; nonhuman; priority journal; structure analysis; Acetobacter; Amino Acids, Essential; Catalysis; Mannosyltransferases; Nuclear Magnetic Resonance, Biomolecular; Protein Structure, Secondary; Recombinant Proteins; Acetobacter; Bacteria (microorganisms); Escherichia coli; Gluconacetobacter xylinus; Xanthomonas campestris
Año:2000
Volumen:275
Número:51
Página de inicio:40568
Página de fin:40575
DOI: http://dx.doi.org/10.1074/jbc.M007496200
Título revista:Journal of Biological Chemistry
Título revista abreviado:J. Biol. Chem.
ISSN:00219258
CODEN:JBCHA
CAS:Amino Acids, Essential; GDP-mannose - cellobiosyl-diphosphopolyprenol mannosyltransferase, EC 2.4.1.-; Mannosyltransferases, EC 2.4.1.-; Recombinant Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00219258_v275_n51_p40568_Abdian

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Citas:

---------- APA ----------
Abdian, P.L., Lellouch, A.C., Gautier, C., Ielpi, L. & Geremia, R.A. (2000) . Identification of essential amino acids in the bacterial α-mannosyltransferase AceA. Journal of Biological Chemistry, 275(51), 40568-40575.
http://dx.doi.org/10.1074/jbc.M007496200
---------- CHICAGO ----------
Abdian, P.L., Lellouch, A.C., Gautier, C., Ielpi, L., Geremia, R.A. "Identification of essential amino acids in the bacterial α-mannosyltransferase AceA" . Journal of Biological Chemistry 275, no. 51 (2000) : 40568-40575.
http://dx.doi.org/10.1074/jbc.M007496200
---------- MLA ----------
Abdian, P.L., Lellouch, A.C., Gautier, C., Ielpi, L., Geremia, R.A. "Identification of essential amino acids in the bacterial α-mannosyltransferase AceA" . Journal of Biological Chemistry, vol. 275, no. 51, 2000, pp. 40568-40575.
http://dx.doi.org/10.1074/jbc.M007496200
---------- VANCOUVER ----------
Abdian, P.L., Lellouch, A.C., Gautier, C., Ielpi, L., Geremia, R.A. Identification of essential amino acids in the bacterial α-mannosyltransferase AceA. J. Biol. Chem. 2000;275(51):40568-40575.
http://dx.doi.org/10.1074/jbc.M007496200