Pseudomonas aeruginosa PA1006 Is a Persulfide-Modified Protein That Is Critical for Molybdenum Homeostasis

Autor: Thomas H. Darrah, Alan E. Friedman, Barbara H. Iglewski, Michael Maguire, Melanie J. Filiatrault, Johanna M. Schwingel, Nadine E. Van Alst, Gregory Tombline, John D. Lapek
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Proteomics
Models
Molecular
Mutant
lcsh:Medicine
Nitrogen Metabolism
medicine.disease_cause
Biochemistry
Nitrate Reductase
Mass Spectrometry
chemistry.chemical_compound
Protein-fragment complementation assay
Homeostasis
lcsh:Science
chemistry.chemical_classification
0303 health sciences
Mutation
Multidisciplinary
Spectrometric Identification of Proteins
Fourier Analysis
Chemistry
Pseudomonas aeruginosa
Metabolic Pathways
Molybdenum cofactor
Research Article
Molecular Sequence Data
Biology
Sulfides
Nitrate reductase
Peptide Mapping
Inorganic Chemistry
03 medical and health sciences
Biosynthesis
Bacterial Proteins
Chemical Biology
medicine
Animals
Amino Acid Sequence
Protein Interactions
Bioinorganic Chemistry
030304 developmental biology
Molybdenum
Sequence Homology
Amino Acid
030306 microbiology
Cofactors
lcsh:R
Molecular biology
Enzyme
Metabolism
chemistry
Membrane protein
Mutagenesis
lcsh:Q
Zdroj: PLoS ONE
PLoS ONE, Vol 8, Iss 2, p e55593 (2013)
ISSN: 1932-6203
Popis: A companion manuscript revealed that deletion of the Pseudomonas aeruginosa (Pae) PA1006 gene caused pleiotropic defects in metabolism including a loss of all nitrate reductase activities, biofilm maturation, and virulence. Herein, several complementary approaches indicate that PA1006 protein serves as a persulfide-modified protein that is critical for molybdenum homeostasis in Pae. Mutation of a highly conserved Cys22 to Ala or Ser resulted in a loss of PA1006 activity. Yeast-two-hybrid and a green-fluorescent protein fragment complementation assay (GFP-PFCA) in Pae itself revealed that PA1006 interacts with Pae PA3667/CsdA and PA3814/IscS Cys desulfurase enzymes. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) “top-down” analysis of PA1006 purified from Pae revealed that conserved Cys22 is post-translationally modified in vivo in the form a persulfide. Inductively-coupled-plasma (ICP)-MS analysis of ΔPA1006 mutant extracts revealed that the mutant cells contain significantly reduced levels of molybdenum compared to wild-type. GFP-PFCA also revealed that PA1006 interacts with several molybdenum cofactor (MoCo) biosynthesis proteins as well as nitrate reductase maturation factor NarJ and component NarH. These data indicate that a loss of PA1006 protein’s persulfide sulfur and a reduced availability of molybdenum contribute to the phenotype of a ΔPA1006 mutant.
Databáze: OpenAIRE
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