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Crystal structures of the ATPase domains of four human Hsp70 isoforms: HSPA1L/Hsp70-hom, HSPA2/Hsp70-2, HSPA6/Hsp70B', and HSPA5/BiP/GRP78.

Wisniewska M, Karlberg T, Lehtiö L, Johansson I, Kotenyova T, Moche M, Schüler H - PLoS ONE (2010)

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Affiliation: Structural Genomics Consortium, Karolinska Institutet, Stockholm, Sweden.

ABSTRACT

Unlabelled: The 70-kDa heat shock proteins (Hsp70) are chaperones with central roles in processes that involve polypeptide remodeling events. Hsp70 proteins consist of two major functional domains: an N-terminal nucleotide binding domain (NBD) with ATPase activity, and a C-terminal substrate binding domain (SBD). We present the first crystal structures of four human Hsp70 isoforms, those of the NBDs of HSPA1L, HSPA2, HSPA5 and HSPA6. As previously with Hsp70 family members, all four proteins crystallized in a closed cleft conformation, although a slight cleft opening through rotation of subdomain IIB was observed for the HSPA5-ADP complex. The structures presented here support the view that the NBDs of human Hsp70 function by conserved mechanisms and contribute little to isoform specificity, which instead is brought about by the SBDs and by accessory proteins.

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Relationship between the ATPase domains of human Hsp70 isoforms.Sequence alignment of the NBDs of selected Hsp70 proteins. Secondary structure elements are indicated for HSPA1A above and for HSPA6 below the alignment. Sequences shown are human HSPA1A (1HJO; gi:5123454); HSPA1L (3GDQ; gi:124256496); HSPA2 (3I33; gi:13676857); bovine Hsc70 (PDB entry 1YUW; gi:76253709), E.coli DnaK (1DKG; gi:16128008); HSPA5 (3IUC; gi:16507237); HSPA9 (no structure available; gi:24234688); and HSPA6 (3FE1; gi:34419635).
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pone-0008625-g001: Relationship between the ATPase domains of human Hsp70 isoforms.Sequence alignment of the NBDs of selected Hsp70 proteins. Secondary structure elements are indicated for HSPA1A above and for HSPA6 below the alignment. Sequences shown are human HSPA1A (1HJO; gi:5123454); HSPA1L (3GDQ; gi:124256496); HSPA2 (3I33; gi:13676857); bovine Hsc70 (PDB entry 1YUW; gi:76253709), E.coli DnaK (1DKG; gi:16128008); HSPA5 (3IUC; gi:16507237); HSPA9 (no structure available; gi:24234688); and HSPA6 (3FE1; gi:34419635).

Mentions: A common feature of Hsp70 ATPase domains is that, despite the large conformational changes they are predicted to undergo in their physiological context, their crystal structures are highly similar under different nucleotide conditions. We addressed the question whether this is also true for the NBDs of HSPA1L, HSPA2, HSPA5 and HSPA6. These share between 67 and 92% sequence identity with the NBD of the major stress inducible isoform, HSPA1A (Figure 1). We used a multiconstruct approach [37] to produce the NBDs of these human Hsp70 isoforms for structure determination by X-ray crystallography. We also produced the NBD of HSPA1A, the structure of which has been determined previously [38], [39]. All five proteins were straightforward to produce in E. coli, although the soluble expression levels for HSPA5/BiP were relatively low.


Crystal structures of the ATPase domains of four human Hsp70 isoforms: HSPA1L/Hsp70-hom, HSPA2/Hsp70-2, HSPA6/Hsp70B', and HSPA5/BiP/GRP78.

Wisniewska M, Karlberg T, Lehtiö L, Johansson I, Kotenyova T, Moche M, Schüler H - PLoS ONE (2010)

Relationship between the ATPase domains of human Hsp70 isoforms.Sequence alignment of the NBDs of selected Hsp70 proteins. Secondary structure elements are indicated for HSPA1A above and for HSPA6 below the alignment. Sequences shown are human HSPA1A (1HJO; gi:5123454); HSPA1L (3GDQ; gi:124256496); HSPA2 (3I33; gi:13676857); bovine Hsc70 (PDB entry 1YUW; gi:76253709), E.coli DnaK (1DKG; gi:16128008); HSPA5 (3IUC; gi:16507237); HSPA9 (no structure available; gi:24234688); and HSPA6 (3FE1; gi:34419635).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2803158&req=5

pone-0008625-g001: Relationship between the ATPase domains of human Hsp70 isoforms.Sequence alignment of the NBDs of selected Hsp70 proteins. Secondary structure elements are indicated for HSPA1A above and for HSPA6 below the alignment. Sequences shown are human HSPA1A (1HJO; gi:5123454); HSPA1L (3GDQ; gi:124256496); HSPA2 (3I33; gi:13676857); bovine Hsc70 (PDB entry 1YUW; gi:76253709), E.coli DnaK (1DKG; gi:16128008); HSPA5 (3IUC; gi:16507237); HSPA9 (no structure available; gi:24234688); and HSPA6 (3FE1; gi:34419635).
Mentions: A common feature of Hsp70 ATPase domains is that, despite the large conformational changes they are predicted to undergo in their physiological context, their crystal structures are highly similar under different nucleotide conditions. We addressed the question whether this is also true for the NBDs of HSPA1L, HSPA2, HSPA5 and HSPA6. These share between 67 and 92% sequence identity with the NBD of the major stress inducible isoform, HSPA1A (Figure 1). We used a multiconstruct approach [37] to produce the NBDs of these human Hsp70 isoforms for structure determination by X-ray crystallography. We also produced the NBD of HSPA1A, the structure of which has been determined previously [38], [39]. All five proteins were straightforward to produce in E. coli, although the soluble expression levels for HSPA5/BiP were relatively low.

Bottom Line: This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions.Please note that a web plugin is required to access this enhanced functionality.Instructions for the installation and use of the web plugin are available in Text S1.

View Article: PubMed Central - PubMed

Affiliation: Structural Genomics Consortium, Karolinska Institutet, Stockholm, Sweden.

ABSTRACT

Unlabelled: The 70-kDa heat shock proteins (Hsp70) are chaperones with central roles in processes that involve polypeptide remodeling events. Hsp70 proteins consist of two major functional domains: an N-terminal nucleotide binding domain (NBD) with ATPase activity, and a C-terminal substrate binding domain (SBD). We present the first crystal structures of four human Hsp70 isoforms, those of the NBDs of HSPA1L, HSPA2, HSPA5 and HSPA6. As previously with Hsp70 family members, all four proteins crystallized in a closed cleft conformation, although a slight cleft opening through rotation of subdomain IIB was observed for the HSPA5-ADP complex. The structures presented here support the view that the NBDs of human Hsp70 function by conserved mechanisms and contribute little to isoform specificity, which instead is brought about by the SBDs and by accessory proteins.

Enhanced version: This article can also be viewed as an enhanced version in which the text of the article is integrated with interactive 3D representations and animated transitions. Please note that a web plugin is required to access this enhanced functionality. Instructions for the installation and use of the web plugin are available in Text S1.

Show MeSH
Related in: MedlinePlus