Limits...
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)

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

Purification and crystallization of Hsp70 isoforms.(A) Coomassie-stained SDS-polyacrylamide gel showing the purity of the crystallized proteins. (B-F) Examples of crystals grown under the conditions that yielded the datasets. (B) HSPA1A; (C) HSPA1L; (D) HSPA2; (E) HSPA5; (F) HSPA6.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2803158&req=5

pone-0008625-g002: Purification and crystallization of Hsp70 isoforms.(A) Coomassie-stained SDS-polyacrylamide gel showing the purity of the crystallized proteins. (B-F) Examples of crystals grown under the conditions that yielded the datasets. (B) HSPA1A; (C) HSPA1L; (D) HSPA2; (E) HSPA5; (F) HSPA6.

Mentions: Crystals of the five proteins were obtained with NBD constructs that contained either the full length N-terminus or short N-terminal truncations and with ADP and either Mg2+- or Mn2+-ions present in the crystallization solution (Figure 2 and Table 1). Crystals diffracted to a resolution of between 1.8 and 2.2 Å except for the HSPA2 crystals which diffracted to 1.3 Å. The structures were solved by molecular replacement (Table 2). In all ATPase domains except for that of HSPA5/BiP the products of ATP hydrolysis, including inorganic phosphate, were observed. Examples of the electron density around the bound nucleotide are shown for HSPA2 and HSPA5 (Figure 3).


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)

Purification and crystallization of Hsp70 isoforms.(A) Coomassie-stained SDS-polyacrylamide gel showing the purity of the crystallized proteins. (B-F) Examples of crystals grown under the conditions that yielded the datasets. (B) HSPA1A; (C) HSPA1L; (D) HSPA2; (E) HSPA5; (F) HSPA6.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2803158&req=5

pone-0008625-g002: Purification and crystallization of Hsp70 isoforms.(A) Coomassie-stained SDS-polyacrylamide gel showing the purity of the crystallized proteins. (B-F) Examples of crystals grown under the conditions that yielded the datasets. (B) HSPA1A; (C) HSPA1L; (D) HSPA2; (E) HSPA5; (F) HSPA6.
Mentions: Crystals of the five proteins were obtained with NBD constructs that contained either the full length N-terminus or short N-terminal truncations and with ADP and either Mg2+- or Mn2+-ions present in the crystallization solution (Figure 2 and Table 1). Crystals diffracted to a resolution of between 1.8 and 2.2 Å except for the HSPA2 crystals which diffracted to 1.3 Å. The structures were solved by molecular replacement (Table 2). In all ATPase domains except for that of HSPA5/BiP the products of ATP hydrolysis, including inorganic phosphate, were observed. Examples of the electron density around the bound nucleotide are shown for HSPA2 and HSPA5 (Figure 3).

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