Dissection of the regulatory role for the N-terminal domain in Candida albicans protein phosphatase Z1

Krisztina Szabó, Zoltán Kónya, F. Erdődi, Ilona Farkas, V. Dombrádi

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

The novel type, fungus specific protein phosphatase Z1 of the opportunistic pathogen, Candida albicans (CaPpz1) has several important physiological roles. It consists of a conserved C-terminal catalytic domain and a variable, intrinsically disordered, N-terminal regulatory domain. To test the function of these domains we modified the structure of CaPpz1 by in vitro mutagenesis. The two main domains were separated, four potential protein binding regions were deleted, and the myristoylation site as well as the active site of the enzyme was crippled by point mutations G2A and R262L, respectively. The in vitro phosphatase activity assay of the bacterially expressed recombinant proteins indicated that the N-terminal domain was inactive, while the C-terminal domain became highly active against myosin light chain substrate. The deletion of the N-terminal 1-16 amino acids and the G2A mutation significantly decreased the specific activity of the enzyme. Complementation of the ppz1 Saccharomyces cerevisiae deletion mutant strain with the different CaPpz1 forms demonstrated that the scission of the main domains, the two point mutations and the N-terminal 1-16 deletion rendered the phosphatase incompetent in the in vivo assays of LiCl tolerance and caffeine sensitivity. Thus our results confirmed the functional role of the N-terminal domain and highlighted the significance of the very N-terminal part of the protein in the regulation of CaPpz1.

Original languageEnglish
Article numbere0211426
JournalPLoS One
Volume14
Issue number2
DOIs
Publication statusPublished - Feb 1 2019

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Dissection
Candida
Phosphoprotein Phosphatases
point mutation
Candida albicans
Phosphoric Monoester Hydrolases
Point Mutation
active sites
Assays
Catalytic Domain
myosin light chains
Myosin Light Chains
Mutagenesis
protein binding
assays
Pathogens
Enzymes
caffeine
Caffeine
Fungi

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Dissection of the regulatory role for the N-terminal domain in Candida albicans protein phosphatase Z1. / Szabó, Krisztina; Kónya, Zoltán; Erdődi, F.; Farkas, Ilona; Dombrádi, V.

In: PLoS One, Vol. 14, No. 2, e0211426, 01.02.2019.

Research output: Contribution to journalArticle

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