Conformational properties of bacterial DnaK and yeast mitochondrial Hsp70: Role of the divergent C-terminal α-helical subdomain

Fernando Moro, Vanesa Fernández-Sáiz, Olga Slutsky, Abdussalam Azem, Arturo Muga

Research output: Contribution to journalArticlepeer-review

Abstract

Among the eukaryotic members of the Hsp70 family, mitochondrial Hsp70 shows the highest degree of sequence identity with bacterial DnaK. Although they share a functional mechanism and homologous co-chaperones, they are highly specific and cannot be exchanged between Escherichia coli and yeast mitochondria. To provide a structural basis for this finding, we characterized both proteins, as well as two DnaK/mtHsp70 chimeras constructed by domain swapping, using biochemical and biophysical methods. Here, we show that DnaK and mtHsp70 display different conformational and biochemical properties. Replacing different regions of the DnaK peptide-binding domain with those of mtHsp70 results in chimeric proteins that: (a) are not able to support growth of an E. coli DnaK deletion strain at stress temperatures (e.g. 42 °C); (b) show increased accessibility and decreased thermal stability of the peptide-binding pocket; and (c) have reduced activation by bacterial, but not mitochondrial co-chaperones, as compared with DnaK. Importantly, swapping the C-terminal α-helical subdomain promotes a conformational change in the chimeras to an mtHsp70-like conformation. Thus, interaction with bacterial co-chaperones correlates well with the conformation that natural and chimeric Hsp70s adopt in solution. Our results support the hypothesis that a specific protein structure might regulate the interaction of Hsp70s with particular components of the cellular machinery, such as Tim44, so that they perform specific functions.

Original languageEnglish
Pages (from-to)3184-3196
Number of pages13
JournalFEBS Journal
Volume272
Issue number12
DOIs
StatePublished - Jun 2005

Keywords

  • Allosterism
  • Chaperones
  • DnaK
  • Mitochondrial Hsp70

Fingerprint

Dive into the research topics of 'Conformational properties of bacterial DnaK and yeast mitochondrial Hsp70: Role of the divergent C-terminal α-helical subdomain'. Together they form a unique fingerprint.

Cite this