skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Uncovering the transmembrane metal binding site of the novel bacterial major facilitator superfamily-type copper importer CcoA

Journal Article · · mBio (Online)
 [1];  [1];  [2];  [1]
  1. Univ. of Pennsylvania, Philadelphia, PA (United States)
  2. Albert-Ludwigs Univ. of Freiburg, Freiburg (Germany)
+ Show Author Affiliations

In this study, uptake and trafficking of metals and their delivery to their respective metalloproteins are important processes. Cells need precise control of each step to avoid exposure to excessive metal concentrations and their harmful consequences. Copper (Cu) is a required micronutrient used as a cofactor in proteins. However, in large amounts, it can induce oxidative damage; hence, Cu homeostasis is indispensable for cell survival. Biogenesis of respiratory heme-Cu oxygen (HCO) reductases includes insertion of Cu into their catalytic subunits to form heme-Cu binuclear centers. Previously, we had shown that CcoA is a major facilitator superfamily (MFS)-type bacterial Cu importer required for biogenesis of cbb3-type cytochromecoxidase (cbb3-Cox). Here, using Rhodobacter capsulatus, we focused on the import and delivery of Cu to cbb3-Cox. By comparing the CcoA amino acid sequence with its homologues from other bacterial species, we located several well-conserved Met, His, and Tyr residues that might be important for Cu transport. We determined the topology of the transmembrane helices that carry these residues to establish that they are membrane embedded, and substituted for them amino acids that do not ligand metal atoms. Characterization of these mutants for their uptake of radioactive64Cu and cbb3-Cox activities demonstrated that Met233 and His261 of CcoA are essential and Met237 and Met265 are important, whereas Tyr230 has no role for Cu uptake or cbb3-Cox biogenesis. These findings show for the first time that CcoA-mediated Cu import relies on conserved Met and His residues that could act as metal ligands at the membrane-embedded Cu binding domain of this transporter.

Research Organization:
Univ. of Pennsylvania, Philadelphia, PA (United States)
Sponsoring Organization:
USDOE Office of Science (SC), Basic Energy Sciences (BES)
Grant/Contract Number:
FG02-91ER20052
OSTI ID:
1258013
Journal Information:
mBio (Online), Vol. 7, Issue 1; Related Information: http://mbio.asm.org/lookup/suppl/doi:10.1128/mBio.01981-15/-/DCSupplemental; ISSN 2150-7511
Publisher:
American Society for MicrobiologyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 12 works
Citation information provided by
Web of Science

References (52)

How copper traverses cellular membranes through the mammalian copper transporter 1, Ctr1: Mammalian copper uptake and mobilization by Ctr1 journal February 2014
Analysis of the regulation of Escherichia coli alkaline phosphatase synthesis using deletions and φ80 transducing phages journal August 1975
Molecular Analysis of the Copper-Transporting Efflux System CusCFBA of Escherichia coli journal July 2003
Involvement of SenC in Assembly of Cytochrome c Oxidase in Rhodobacter capsulatus journal November 2005
The Mechanism of Copper Uptake Mediated by Human CTR1: A MUTATIONAL ANALYSIS journal August 2005
Export of the siderophore enterobactin in Escherichia coli: involvement of a 43 kDa membrane exporter: Enterobactin secretion journal May 2002
The structure and function of heavy metal transport P1B-ATPases journal January 2007
Mechanisms of copper homeostasis in bacteria journal January 2013
The ScoI homologue SenC is a copper binding protein that interacts directly with the cbb3-type cytochrome oxidase in Rhodobacter capsulatus journal November 2012
Copper trafficking to the mitochondrion and assembly of copper metalloenzymes journal July 2006
Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 journal August 1970
Isolation and Characterization of Rhodobacter capsulatus Mutants Affected in Cytochromecbb 3 Oxidase Activity journal January 1998
Crystal structures of the CusA efflux pump suggest methionine-mediated metal transport journal September 2010
Mechanisms for copper acquisition, distribution and regulation journal February 2008
Biogenesis of cbb3-type cytochrome c oxidase in Rhodobacter capsulatus journal June 2012
Structural advances for the major facilitator superfamily (MFS) transporters journal March 2013
Understanding Copper Trafficking in Bacteria:  Interaction between the Copper Transport Protein CopZ and the N-Terminal Domain of the Copper ATPase CopA from Bacillus subtilis † journal February 2003
Structure of the Multidrug Transporter EmrD from Escherichia coli journal May 2006
Interactions between CusF and CusB Identified by NMR Spectroscopy and Chemical Cross-Linking Coupled to Mass Spectrometry journal April 2011
CtpA, a Copper-translocating P-type ATPase Involved in the Biogenesis of Multiple Copper-requiring Enzymes journal April 2010
Molecular characterization of a copper transport protein in S. cerevisiae: An unexpected role for copper in iron transport journal January 1994
A redox switch in CopC: An intriguing copper trafficking protein that binds copper(I) and copper(II) at different sites journal March 2003
Intracytoplasmic Copper Homeostasis Controls Cytochrome c Oxidase Production journal January 2014
Copper Metallochaperones journal June 2010
Determinants of membrane protein topology. journal December 1987
Crystal structure of a copper-transporting PIB-type ATPase journal June 2011
The Major Facilitator Superfamily-Type Transporter YmfE and the Multidrug-Efflux Activator Mta Mediate Bacillibactin Secretion in Bacillus subtilis journal May 2008
Novel Rhodobacter capsulatus genes required for the biogenesis of various c-type cytochromes journal January 2000
Novel Transporter Required for Biogenesis of cbb3-Type Cytochrome c Oxidase in Rhodobacter capsulatus journal January 2012
Three-dimensional structure of the human copper transporter hCTR1 journal February 2009
Nutritional immunity: transition metals at the pathogen–host interface journal July 2012
Projection structure of the human copper transporter CTR1 at 6-A resolution reveals a compact trimer with a novel channel-like architecture journal February 2006
The Structure of cbb3 Cytochrome Oxidase Provides Insights into Proton Pumping journal June 2010
Human copper homeostasis: a network of interconnected pathways journal April 2010
The [2Fe-2S] Cluster E m as an Indicator of the Iron-Sulfur Subunit Position in the Ubihydroquinone Oxidation Site of the Cytochrome bc 1 Complex journal November 2001
The P-Type ATPase Superfamily journal January 2010
Crystal structure of a bacterial homologue of glucose transporters GLUT1–4 journal October 2012
Structure and Mechanism of the Lactose Permease of Escherichia coli journal August 2003
Distinct functional roles of homologous Cu+ efflux ATPases in Pseudomonas aeruginosa: Role of Cu+ transport ATPases in Pseudomonas aeruginosa journal October 2010
P-Type ATPases journal June 2011
Structure of the YajR transporter suggests a transport mechanism based on the conserved motif A journal August 2013
Prokaryotic Cu,Zn superoxide dismutases journal December 2003
Mechanism of Cu+-transporting ATPases: Soluble Cu+ chaperones directly transfer Cu+ to transmembrane transport sites journal April 2008
Coordination chemistry of copper proteins: How nature handles a toxic cargo for essential function journal February 2012
A Novel Copper-Binding Fold for the Periplasmic Copper Resistance Protein CusF † , ‡ journal August 2005
Introduction to Human Disorders of Copper Metabolism: Introduction journal May 2014
Biochemical and Genetic Analyses of Yeast and Human High Affinity Copper Transporters Suggest a Conserved Mechanism for Copper Uptake journal April 2002
Laccases of prokaryotic origin: enzymes at the interface of protein science and protein technology journal January 2015
Copper Transporting P-Type ATPases and Human Disease journal January 2002
Altered microglial copper homeostasis in a mouse model of Alzheimer’s disease: ATP7A protein expression in AD mouse brain journal September 2010
Roles of the ccoGHIS gene products in the biogenesis of the cbb3-type cytochrome c oxidase journal March 2000
Crystal structure of the CusBA heavy-metal efflux complex of Escherichia coli journal February 2011

Cited By (2)

The cbb 3 -type cytochrome oxidase assembly factor CcoG is a widely distributed cupric reductase journal September 2019
Cu Transport by the Extended Family of CcoA-like Transporters (CalT) in Proteobacteria journal February 2019

Similar Records

Cysteine Mutants of the Major Facilitator Superfamily-Type Transporter CcoA Provide Insight into Copper Import
Journal Article · Tue Aug 31 00:00:00 EDT 2021 · mBio (Online) · OSTI ID:1258013
+3 more
Widespread Distribution and Functional Specificity of the Copper Importer CcoA: Distinct Cu Uptake Routes for Bacterial Cytochrome c Oxidases
Journal Article · Tue Feb 27 00:00:00 EST 2018 · mBio (Online) · OSTI ID:1258013
+5 more
Cu Transport by the Extended Family of CcoA-like Transporters (CalT) in Proteobacteria
Journal Article · Mon Feb 04 00:00:00 EST 2019 · Scientific Reports · OSTI ID:1258013
+5 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES