Biochemical and genetic analyses of yeast and human high affinity copper transporters suggest a conserved mechanism for copper uptake

Sergi Puig, Jaekwon Lee, Miranda Lau, Dennis J. Thiele

Research output: Contribution to journalArticle

273 Citations (Scopus)

Abstract

The redox active metal copper is an essential cofactor in critical biological processes such as respiration, iron transport, oxidative stress protection, hormone production, and pigmentation. A widely conserved family of high affinity copper transport proteins (Ctr proteins) mediates copper uptake at the plasma membrane. However, little is known about Ctr protein topology, structure, and the mechanisms by which this class of transporters mediates high affinity copper uptake. In this report, we elucidate the topological orientation of the yeast Ctr1 copper transport protein. We show that a series of clustered methionine residues in the hydrophilic extracellular domain and an MXXXM motif in the second transmembrane domain are important for copper uptake but not for protein sorting and delivery to the cell surface. The conversion of these methionine residues to cysteine, by site-directed mutagenesis, strongly suggests that they coordinate to copper during the process of metal transport. Genetic evidence supports an essential role for cooperativity between monomers for the formation of an active Ctr transport complex. Together, these results support a fundamentally conserved mechanism for high affinity copper uptake through the Ctr proteins in yeast and humans.

Original languageEnglish (US)
Pages (from-to)26021-26030
Number of pages10
JournalJournal of Biological Chemistry
Volume277
Issue number29
DOIs
StatePublished - Jul 19 2002

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Yeast
Copper
Molecular Biology
Yeasts
Carrier Proteins
Methionine
Metals
Biological Phenomena
Mutagenesis
Oxidative stress
Active Biological Transport
Pigmentation
Protein Transport
Cell membranes
Site-Directed Mutagenesis
Sorting
Oxidation-Reduction
Cysteine
Respiration
Proteins

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Biochemical and genetic analyses of yeast and human high affinity copper transporters suggest a conserved mechanism for copper uptake. / Puig, Sergi; Lee, Jaekwon; Lau, Miranda; Thiele, Dennis J.

In: Journal of Biological Chemistry, Vol. 277, No. 29, 19.07.2002, p. 26021-26030.

Research output: Contribution to journalArticle

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