October 16, 2012

The Hsp70 family is ubiquitous

  • Hsp70 is a chaperone that functions on target proteins in conjunction with DnaJ and GrpE.
  • Members of the Hsp70 family are found in the cytosol, in the ER, and in mitochondria and chloroplasts.
The Hsp70 family is found in bacteria, eukaryotic cytosol, in the endoplasmic reticulum, and in chloroplasts and mitochondria (for review see Frydman, 2001). A typical Hsp70 has two domains: the N-terminal domain is an ATPase; and the C-terminal domain binds the substrate polypeptide (Flaherty, DeLuca-Flaherty, and McKay, 1990; Zhu et al., 1996). When bound to ATP, Hsp70 binds and releases substrates rapidly; when bound to ADP, the reactions are slow. Recycling between these states is regulated by two other proteins, Hsp40 (DnaJ) and GrpE.

Figure 8.13 shows that Hsp40 (DnaJ) binds first to a nascent protein as it emerges from the ribosome. Hsp40 contains a region called the J domain (named for DnaJ), which interacts with Hsp70. Hsp70 (DnaK) binds to both Hsp40 and to the unfolded protein. In effect, two interacting chaperones bind to the protein. The J domain accounts for the specificity of the pairwise interaction, and drives a particular Hsp40 to select the appropriate partner from the Hsp70 family.
The interaction of Hsp70 (DnaK) with Hsp40 (DnaJ) stimulates the ATPase activity of Hsp70. The ADP-bound form of the complex remains associated with the protein substrate until GrpE displaces the ADP. This causes loss of Hsp40 followed by dissociation of Hsp70. The Hsp70 binds another ATP and the cycle can be repeated. GrpE (or its equivalent) is found only in bacteria, mitochondria, and chloroplasts; in other locations, the dissociation reaction is coupled to ATP hydrolysis in a more complex way.
Protein folding is accomplished by multiple cycles of association and dissociation. As the protein chain lengthens, Hsp70 (DnaK) may dissociate from one binding site and then reassociate with another, thus releasing parts of the substrate protein to fold correctly in an ordered manner. Finally, the intact protein is released from the ribosome, folded into its mature conformation (for review see Georgopoulos and Welch, 1993; Hartl, 1966; Bukau and Horwich, 1998).
Different members of the Hsp70 class function on various types of target proteins. Cytosolic proteins (the eponymous Hsp70 and a related protein called Hsc70) act on nascent proteins on ribosomes. Variants in the ER (called BiP or Grp78 in higher eukaryotes, called Kar2 in S. cerevisiae), or in mitochondria or chloroplasts, function in a rather similar manner on proteins as they emerge into the interior of the organelle on passing through the membrane.
What feature does Hsp70 recognize in a target protein? It binds to a linear stretch of amino acids embedded in a hydrophobic context (Flynn et al., 1991; Blond-Elguindi et al., 1993). This is precisely the sort of motif that is buried in the hydrophobic core of a properly folded, mature protein. Its exposure therefore indicates that the protein is nascent or denatured. Motifs of this nature occur about every 40 amino acids. Binding to the motif prevents it from misaggregating with another one.
This mode of action explains how the Hsp70 protein Bip can fulfill two functions: to assist in oligomerization and/or folding of newly translocated proteins in the ER; and to remove misfolded proteins. Suppose that BiP recognizes certain peptide sequences that are inaccessible within the conformation of a mature, properly folded protein. These sequences are exposed and attract BiP when the protein enters the ER lumen in an essentially one-dimensional form. And if a protein is misfolded or denatured, they may become exposed on its surface instead of being properly buried.