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Chaperones of F1-ATPase.

Ludlam A, Brunzelle J, Pribyl T, Xu X, Gatti DL, Ackerman SH - J. Biol. Chem. (2009)

Bottom Line: One important feature of this model was the prediction that as long as Atp11p is bound to beta and Atp12p is bound to alpha, the two F(1) subunits cannot interact at either the catalytic site or the noncatalytic site interface.Here we present the structures of Atp11p from Candida glabrata and Atp12p from Paracoccus denitrificans, and we show that some features of the Wang model are correct, namely that binding of the chaperones to alpha and beta prevents further interactions between these F(1) subunits.However, Atp11p and Atp12p do not resemble alpha or beta, and it is instead the F(1) gamma subunit that initiates the release of the chaperones from alpha and beta and their further assembly into the mature complex.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.

ABSTRACT
Mitochondrial F(1)-ATPase contains a hexamer of alternating alpha and beta subunits. The assembly of this structure requires two specialized chaperones, Atp11p and Atp12p, that bind transiently to beta and alpha. In the absence of Atp11p and Atp12p, the hexamer is not formed, and alpha and beta precipitate as large insoluble aggregates. An early model for the mechanism of chaperone-mediated F(1) assembly (Wang, Z. G., Sheluho, D., Gatti, D. L., and Ackerman, S. H. (2000) EMBO J. 19, 1486-1493) hypothesized that the chaperones themselves look very much like the alpha and beta subunits, and proposed an exchange of Atp11p for alpha and of Atp12p for beta; the driving force for the exchange was expected to be a higher affinity of alpha and beta for each other than for the respective chaperone partners. One important feature of this model was the prediction that as long as Atp11p is bound to beta and Atp12p is bound to alpha, the two F(1) subunits cannot interact at either the catalytic site or the noncatalytic site interface. Here we present the structures of Atp11p from Candida glabrata and Atp12p from Paracoccus denitrificans, and we show that some features of the Wang model are correct, namely that binding of the chaperones to alpha and beta prevents further interactions between these F(1) subunits. However, Atp11p and Atp12p do not resemble alpha or beta, and it is instead the F(1) gamma subunit that initiates the release of the chaperones from alpha and beta and their further assembly into the mature complex.

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Pairwise alignment of recombinant protein sequences with S. cerevisiae homologs. Upper, pairwise alignment of C. glabrata and S. cerevisiae Atp11p. The disordered regions of the C. glabrata structure (residues 1–93 and 163–176) are shown in orange, the N-terminal helical domain (residues 94–129) in blue, the central α/β taco (residues 130–262) in green, and the C-terminal helical domain (residues 263–298) in red. Lower, pairwise alignment of P. denitrificans and S. cerevisiae Atp12p. The smaller N-terminal domain (residues 3–83) and the larger C-terminal domain (residues 84–238) are shown in blue and red, respectively. Trp-57 and Asp-202, corresponding, respectively, to Trp-103 and Glu-289 of the yeast protein, are highlighted in blue and red boxes. C. gla, C. glabrata; P. den, P. denitrificans; S. cer., S. cerevisiae.
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Figure 1: Pairwise alignment of recombinant protein sequences with S. cerevisiae homologs. Upper, pairwise alignment of C. glabrata and S. cerevisiae Atp11p. The disordered regions of the C. glabrata structure (residues 1–93 and 163–176) are shown in orange, the N-terminal helical domain (residues 94–129) in blue, the central α/β taco (residues 130–262) in green, and the C-terminal helical domain (residues 263–298) in red. Lower, pairwise alignment of P. denitrificans and S. cerevisiae Atp12p. The smaller N-terminal domain (residues 3–83) and the larger C-terminal domain (residues 84–238) are shown in blue and red, respectively. Trp-57 and Asp-202, corresponding, respectively, to Trp-103 and Glu-289 of the yeast protein, are highlighted in blue and red boxes. C. gla, C. glabrata; P. den, P. denitrificans; S. cer., S. cerevisiae.

Mentions: Crystals of C. glabrata Atp11p and P. denitrificans Atp12p were obtained after screening several homologous proteins, including those from Homo sapiens, Mus musculus, S. cerevisiae, Candida albicans, Kluveromyces lactis, Rhodobacter capsulatus, and Arabidopsis thaliana. C. glabrata Atp11p and P. denitrificans Atp12p are 79.9 and 44.2% similar and 56.3% and 23.4% identical to the corresponding proteins in S. cerevisiae (Fig. 1). When produced from episomal plasmids C. glabrata Atp11p and P. denitrificans Atp12p rescue the respiratory defect of S. cerevisiae atp11 or atp12 deletion strains (Table 1; Fig. 2), suggesting that the mechanism by which the chaperones facilitate the assembly of F1 is maintained across evolutionary lines.


Chaperones of F1-ATPase.

Ludlam A, Brunzelle J, Pribyl T, Xu X, Gatti DL, Ackerman SH - J. Biol. Chem. (2009)

Pairwise alignment of recombinant protein sequences with S. cerevisiae homologs. Upper, pairwise alignment of C. glabrata and S. cerevisiae Atp11p. The disordered regions of the C. glabrata structure (residues 1–93 and 163–176) are shown in orange, the N-terminal helical domain (residues 94–129) in blue, the central α/β taco (residues 130–262) in green, and the C-terminal helical domain (residues 263–298) in red. Lower, pairwise alignment of P. denitrificans and S. cerevisiae Atp12p. The smaller N-terminal domain (residues 3–83) and the larger C-terminal domain (residues 84–238) are shown in blue and red, respectively. Trp-57 and Asp-202, corresponding, respectively, to Trp-103 and Glu-289 of the yeast protein, are highlighted in blue and red boxes. C. gla, C. glabrata; P. den, P. denitrificans; S. cer., S. cerevisiae.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2719352&req=5

Figure 1: Pairwise alignment of recombinant protein sequences with S. cerevisiae homologs. Upper, pairwise alignment of C. glabrata and S. cerevisiae Atp11p. The disordered regions of the C. glabrata structure (residues 1–93 and 163–176) are shown in orange, the N-terminal helical domain (residues 94–129) in blue, the central α/β taco (residues 130–262) in green, and the C-terminal helical domain (residues 263–298) in red. Lower, pairwise alignment of P. denitrificans and S. cerevisiae Atp12p. The smaller N-terminal domain (residues 3–83) and the larger C-terminal domain (residues 84–238) are shown in blue and red, respectively. Trp-57 and Asp-202, corresponding, respectively, to Trp-103 and Glu-289 of the yeast protein, are highlighted in blue and red boxes. C. gla, C. glabrata; P. den, P. denitrificans; S. cer., S. cerevisiae.
Mentions: Crystals of C. glabrata Atp11p and P. denitrificans Atp12p were obtained after screening several homologous proteins, including those from Homo sapiens, Mus musculus, S. cerevisiae, Candida albicans, Kluveromyces lactis, Rhodobacter capsulatus, and Arabidopsis thaliana. C. glabrata Atp11p and P. denitrificans Atp12p are 79.9 and 44.2% similar and 56.3% and 23.4% identical to the corresponding proteins in S. cerevisiae (Fig. 1). When produced from episomal plasmids C. glabrata Atp11p and P. denitrificans Atp12p rescue the respiratory defect of S. cerevisiae atp11 or atp12 deletion strains (Table 1; Fig. 2), suggesting that the mechanism by which the chaperones facilitate the assembly of F1 is maintained across evolutionary lines.

Bottom Line: One important feature of this model was the prediction that as long as Atp11p is bound to beta and Atp12p is bound to alpha, the two F(1) subunits cannot interact at either the catalytic site or the noncatalytic site interface.Here we present the structures of Atp11p from Candida glabrata and Atp12p from Paracoccus denitrificans, and we show that some features of the Wang model are correct, namely that binding of the chaperones to alpha and beta prevents further interactions between these F(1) subunits.However, Atp11p and Atp12p do not resemble alpha or beta, and it is instead the F(1) gamma subunit that initiates the release of the chaperones from alpha and beta and their further assembly into the mature complex.

View Article: PubMed Central - PubMed

Affiliation: Department of Biochemistry and Molecular Biology, Wayne State University School of Medicine, Detroit, Michigan 48201, USA.

ABSTRACT
Mitochondrial F(1)-ATPase contains a hexamer of alternating alpha and beta subunits. The assembly of this structure requires two specialized chaperones, Atp11p and Atp12p, that bind transiently to beta and alpha. In the absence of Atp11p and Atp12p, the hexamer is not formed, and alpha and beta precipitate as large insoluble aggregates. An early model for the mechanism of chaperone-mediated F(1) assembly (Wang, Z. G., Sheluho, D., Gatti, D. L., and Ackerman, S. H. (2000) EMBO J. 19, 1486-1493) hypothesized that the chaperones themselves look very much like the alpha and beta subunits, and proposed an exchange of Atp11p for alpha and of Atp12p for beta; the driving force for the exchange was expected to be a higher affinity of alpha and beta for each other than for the respective chaperone partners. One important feature of this model was the prediction that as long as Atp11p is bound to beta and Atp12p is bound to alpha, the two F(1) subunits cannot interact at either the catalytic site or the noncatalytic site interface. Here we present the structures of Atp11p from Candida glabrata and Atp12p from Paracoccus denitrificans, and we show that some features of the Wang model are correct, namely that binding of the chaperones to alpha and beta prevents further interactions between these F(1) subunits. However, Atp11p and Atp12p do not resemble alpha or beta, and it is instead the F(1) gamma subunit that initiates the release of the chaperones from alpha and beta and their further assembly into the mature complex.

Show MeSH
Related in: MedlinePlus