title: AztD, a Periplasmic Zinc Metallochaperone to an ATP-binding Cassette (ABC) Transporter System in Paracoccus denitrificans.
abstract: Bacterial ATP-binding cassette (ABC) transporters of transition metals are essential for acquisition of necessary elements from the environment. A large number of Gram-negative bacteria, including human pathogens, have a fourth conserved gene of unknown function adjacent to the canonical permease, ATPase, and solute-binding protein (SBP) genes of the AztABC zinc transporter system. To assess the function of this putative accessory factor (AztD) from Paracoccus denitrificans, we have analyzed its transcriptional regulation, metal binding properties, and interaction with the SBP (AztC). Transcription of the aztD gene is significantly up-regulated under conditions of zinc starvation. Recombinantly expressed AztD purifies with slightly substoichiometric zinc from the periplasm of Escherichia coli and is capable of binding up to three zinc ions with high affinity. Size exclusion chromatography and a simple intrinsic fluorescence assay were used to determine that AztD as isolated is able to transfer bound zinc nearly quantitatively to apo-AztC. Transfer occurs through a direct, associative mechanism that prevents loss of metal to the solvent. These results indicate that AztD is a zinc chaperone to AztC and likely functions to maintain zinc homeostasis through interaction with the AztABC system. This work extends our understanding of periplasmic zinc trafficking and the function of chaperones in this process.
authors.affiliation: From the Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico 88003.
authors.forename: Melody
authors.initials: M
authors.lastname: Handali
authors.affiliation: From the Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico 88003.
authors.forename: Hridindu
authors.initials: H
authors.lastname: Roychowdhury
authors.affiliation: From the Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico 88003.
authors.forename: Durga P
authors.initials: DP
authors.lastname: Neupane
authors.affiliation: From the Department of Chemistry and Biochemistry, New Mexico State University, Las Cruces, New Mexico 88003 etyukl@nmsu.edu.
authors.forename: Erik T
authors.initials: ET
authors.lastname: Yukl
mesh_terms: D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D002942:Circular Dichroism; D018832:Molecular Chaperones / Q000378:metabolism*; D010231:Paracoccus denitrificans / Q000378:metabolism*; D019897:Periplasm / Q000378:metabolism*; D011485:Protein Binding; D013050:Spectrometry, Fluorescence; D015032:Zinc / Q000378:metabolism*
chemical_list: D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; D018832:Molecular Chaperones; D015032:Zinc
keywords: ABC transporter; chaperone; metal homeostasis; metalloprotein; zinc