Biochemical characterization of recombinant Candida albicans mannosyltransferases Mnt1, Mnt2 and Mnt5 reveals new functions in O- and N-mannan biosynthesis

Diana F. Diaz-Jimenez, Hector M. Mora-Montes, Arturo Hernandez-Cervantes, Juan P. Luna-Arias, Neil A. R. Gow, Arturo Flores-Carreon

Research output: Contribution to journalArticle

28 Citations (Scopus)
3 Downloads (Pure)

Abstract

The cell surface of Candida albicans is enriched with highly glycosylated mannoproteins that are involved in the interaction with host tissues. N- and O-glycosylation are post-translational modifications that initiate in the endoplasmic reticulum, and finalize in the Golgi. The KRE2/MNT1 family encode a set of multifunctional mannosyltransferases that participate in O-, N- and phosphomannosylation. In order to gain insights into the substrate specificities of these enzymes, recombinant forms of Mnt1. Mnt2, and Mnt5 were expressed in Pichia pastoris and the enzyme activities characterized. Mnt1 and Mnt2 showed a high specificity for alpha-methylmannoside and alpha 1,2-mannobiose as acceptor substrates. Notably, they also used Saccharomyces cerevisiae O-mannans as acceptors and generated products with more than three mannose residues, suggesting than Mnt1 and Mnt2 could be the mannosyltransferases adding the fourth and fifth mannose residue to the O-mannans in C albicans. Mnt5 only recognized alpha-mannans as acceptor, suggesting that participates in the addition of the second mannose residues to the N-glycan outer chain. (C) 2012 Elsevier Inc. All rights reserved.

Original languageEnglish
Pages (from-to)77-82
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume419
Issue number1
DOIs
Publication statusPublished - 2 Mar 2012

Keywords

  • candida albicans
  • mannosyltransferases
  • glycosylation
  • cell-wall integrity
  • saccharomyces-cerevisiae
  • pichia-pastoris
  • protein mannosyltransferase
  • antifungal resistance
  • virulence
  • alpha-1,2-mannosyltransferase
  • morphogenesis
  • recognition

Cite this

Biochemical characterization of recombinant Candida albicans mannosyltransferases Mnt1, Mnt2 and Mnt5 reveals new functions in O- and N-mannan biosynthesis. / Diaz-Jimenez, Diana F.; Mora-Montes, Hector M.; Hernandez-Cervantes, Arturo; Luna-Arias, Juan P.; Gow, Neil A. R.; Flores-Carreon, Arturo.

In: Biochemical and Biophysical Research Communications, Vol. 419, No. 1, 02.03.2012, p. 77-82.

Research output: Contribution to journalArticle

Diaz-Jimenez, Diana F. ; Mora-Montes, Hector M. ; Hernandez-Cervantes, Arturo ; Luna-Arias, Juan P. ; Gow, Neil A. R. ; Flores-Carreon, Arturo. / Biochemical characterization of recombinant Candida albicans mannosyltransferases Mnt1, Mnt2 and Mnt5 reveals new functions in O- and N-mannan biosynthesis. In: Biochemical and Biophysical Research Communications. 2012 ; Vol. 419, No. 1. pp. 77-82.
@article{3efba929e7204e2a9538bf05b78c6783,
title = "Biochemical characterization of recombinant Candida albicans mannosyltransferases Mnt1, Mnt2 and Mnt5 reveals new functions in O- and N-mannan biosynthesis",
abstract = "The cell surface of Candida albicans is enriched with highly glycosylated mannoproteins that are involved in the interaction with host tissues. N- and O-glycosylation are post-translational modifications that initiate in the endoplasmic reticulum, and finalize in the Golgi. The KRE2/MNT1 family encode a set of multifunctional mannosyltransferases that participate in O-, N- and phosphomannosylation. In order to gain insights into the substrate specificities of these enzymes, recombinant forms of Mnt1. Mnt2, and Mnt5 were expressed in Pichia pastoris and the enzyme activities characterized. Mnt1 and Mnt2 showed a high specificity for alpha-methylmannoside and alpha 1,2-mannobiose as acceptor substrates. Notably, they also used Saccharomyces cerevisiae O-mannans as acceptors and generated products with more than three mannose residues, suggesting than Mnt1 and Mnt2 could be the mannosyltransferases adding the fourth and fifth mannose residue to the O-mannans in C albicans. Mnt5 only recognized alpha-mannans as acceptor, suggesting that participates in the addition of the second mannose residues to the N-glycan outer chain. (C) 2012 Elsevier Inc. All rights reserved.",
keywords = "candida albicans, mannosyltransferases, glycosylation, cell-wall integrity, saccharomyces-cerevisiae, pichia-pastoris, protein mannosyltransferase, antifungal resistance, virulence, alpha-1,2-mannosyltransferase, morphogenesis, recognition",
author = "Diaz-Jimenez, {Diana F.} and Mora-Montes, {Hector M.} and Arturo Hernandez-Cervantes and Luna-Arias, {Juan P.} and Gow, {Neil A. R.} and Arturo Flores-Carreon",
year = "2012",
month = "3",
day = "2",
doi = "10.1016/j.bbrc.2012.01.131",
language = "English",
volume = "419",
pages = "77--82",
journal = "Biochemical and Biophysical Research Communications",
issn = "0006-291X",
publisher = "Academic Press Inc.",
number = "1",

}

TY - JOUR

T1 - Biochemical characterization of recombinant Candida albicans mannosyltransferases Mnt1, Mnt2 and Mnt5 reveals new functions in O- and N-mannan biosynthesis

AU - Diaz-Jimenez, Diana F.

AU - Mora-Montes, Hector M.

AU - Hernandez-Cervantes, Arturo

AU - Luna-Arias, Juan P.

AU - Gow, Neil A. R.

AU - Flores-Carreon, Arturo

PY - 2012/3/2

Y1 - 2012/3/2

N2 - The cell surface of Candida albicans is enriched with highly glycosylated mannoproteins that are involved in the interaction with host tissues. N- and O-glycosylation are post-translational modifications that initiate in the endoplasmic reticulum, and finalize in the Golgi. The KRE2/MNT1 family encode a set of multifunctional mannosyltransferases that participate in O-, N- and phosphomannosylation. In order to gain insights into the substrate specificities of these enzymes, recombinant forms of Mnt1. Mnt2, and Mnt5 were expressed in Pichia pastoris and the enzyme activities characterized. Mnt1 and Mnt2 showed a high specificity for alpha-methylmannoside and alpha 1,2-mannobiose as acceptor substrates. Notably, they also used Saccharomyces cerevisiae O-mannans as acceptors and generated products with more than three mannose residues, suggesting than Mnt1 and Mnt2 could be the mannosyltransferases adding the fourth and fifth mannose residue to the O-mannans in C albicans. Mnt5 only recognized alpha-mannans as acceptor, suggesting that participates in the addition of the second mannose residues to the N-glycan outer chain. (C) 2012 Elsevier Inc. All rights reserved.

AB - The cell surface of Candida albicans is enriched with highly glycosylated mannoproteins that are involved in the interaction with host tissues. N- and O-glycosylation are post-translational modifications that initiate in the endoplasmic reticulum, and finalize in the Golgi. The KRE2/MNT1 family encode a set of multifunctional mannosyltransferases that participate in O-, N- and phosphomannosylation. In order to gain insights into the substrate specificities of these enzymes, recombinant forms of Mnt1. Mnt2, and Mnt5 were expressed in Pichia pastoris and the enzyme activities characterized. Mnt1 and Mnt2 showed a high specificity for alpha-methylmannoside and alpha 1,2-mannobiose as acceptor substrates. Notably, they also used Saccharomyces cerevisiae O-mannans as acceptors and generated products with more than three mannose residues, suggesting than Mnt1 and Mnt2 could be the mannosyltransferases adding the fourth and fifth mannose residue to the O-mannans in C albicans. Mnt5 only recognized alpha-mannans as acceptor, suggesting that participates in the addition of the second mannose residues to the N-glycan outer chain. (C) 2012 Elsevier Inc. All rights reserved.

KW - candida albicans

KW - mannosyltransferases

KW - glycosylation

KW - cell-wall integrity

KW - saccharomyces-cerevisiae

KW - pichia-pastoris

KW - protein mannosyltransferase

KW - antifungal resistance

KW - virulence

KW - alpha-1,2-mannosyltransferase

KW - morphogenesis

KW - recognition

U2 - 10.1016/j.bbrc.2012.01.131

DO - 10.1016/j.bbrc.2012.01.131

M3 - Article

VL - 419

SP - 77

EP - 82

JO - Biochemical and Biophysical Research Communications

JF - Biochemical and Biophysical Research Communications

SN - 0006-291X

IS - 1

ER -