Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase
Abstract
Here, the ring-forming Hsp104 ATPase cooperates with Hsp70 and Hsp40 molecular chaperones to rescue stress-damaged proteins from both amorphous and amyloid-forming aggregates. The ability to do so relies upon pore loops present in the first ATP-binding domain (AAA-1; loop-1 and loop-2) and in the second ATP-binding domain (AAA-2; loop-3) of Hsp104, which face the protein translocating channel and couple ATP-driven changes in pore loop conformation to substrate translocation. A hallmark of loop-1 and loop-3 is an invariable and mutational sensitive aromatic amino acid (Tyr257 and Tyr662) involved in substrate binding. However, the role of conserved aliphatic residues (Lys256, Lys258, and Val663) flanking the pore loop tyrosines, and the function of loop-2 in protein disaggregation has not been investigated. Here we present the crystal structure of an N-terminal fragment of Saccharomyces cerevisiae Hsp104 exhibiting molecular interactions involving both AAA-1 pore loops, which resemble contacts with bound substrate. Corroborated by biochemical experiments and functional studies in yeast, we show that aliphatic residues flanking Tyr257 and Tyr662 are equally important for substrate interaction, and abolish Hsp104 function when mutated to glycine. Unexpectedly, we find that loop-2 is sensitive to aspartate substitutions that impair Hsp104 function and abolish protein disaggregation when loop-2 is replacedmore »
- Authors:
-
- Baylor College of Medicine, Houston, TX (United States)
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Publication Date:
- Research Org.:
- Argonne National Lab. (ANL), Argonne, IL (United States)
- Sponsoring Org.:
- National Institutes of Health (NIH); USDOE Office of Science (SC), Biological and Environmental Research (BER)
- OSTI Identifier:
- 1461438
- Grant/Contract Number:
- AC02-06CH11357
- Resource Type:
- Accepted Manuscript
- Journal Name:
- Bioscience Reports
- Additional Journal Information:
- Journal Volume: 37; Journal Issue: 6; Journal ID: ISSN 0144-8463
- Publisher:
- Portland Press - Biochemical Society
- Country of Publication:
- United States
- Language:
- English
- Subject:
- 59 BASIC BIOLOGICAL SCIENCES
Citation Formats
Lee, Jungsoon, Sung, Nuri, Yeo, Lythou, Chang, Changsoo, Lee, Sukyeong, and Tsai, Francis T. F. Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase. United States: N. p., 2017.
Web. doi:10.1042/BSR20171399.
Lee, Jungsoon, Sung, Nuri, Yeo, Lythou, Chang, Changsoo, Lee, Sukyeong, & Tsai, Francis T. F. Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase. United States. https://doi.org/10.1042/BSR20171399
Lee, Jungsoon, Sung, Nuri, Yeo, Lythou, Chang, Changsoo, Lee, Sukyeong, and Tsai, Francis T. F. Fri .
"Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase". United States. https://doi.org/10.1042/BSR20171399. https://www.osti.gov/servlets/purl/1461438.
@article{osti_1461438,
title = {Structural determinants for protein unfolding and translocation by the Hsp104 protein disaggregase},
author = {Lee, Jungsoon and Sung, Nuri and Yeo, Lythou and Chang, Changsoo and Lee, Sukyeong and Tsai, Francis T. F.},
abstractNote = {Here, the ring-forming Hsp104 ATPase cooperates with Hsp70 and Hsp40 molecular chaperones to rescue stress-damaged proteins from both amorphous and amyloid-forming aggregates. The ability to do so relies upon pore loops present in the first ATP-binding domain (AAA-1; loop-1 and loop-2) and in the second ATP-binding domain (AAA-2; loop-3) of Hsp104, which face the protein translocating channel and couple ATP-driven changes in pore loop conformation to substrate translocation. A hallmark of loop-1 and loop-3 is an invariable and mutational sensitive aromatic amino acid (Tyr257 and Tyr662) involved in substrate binding. However, the role of conserved aliphatic residues (Lys256, Lys258, and Val663) flanking the pore loop tyrosines, and the function of loop-2 in protein disaggregation has not been investigated. Here we present the crystal structure of an N-terminal fragment of Saccharomyces cerevisiae Hsp104 exhibiting molecular interactions involving both AAA-1 pore loops, which resemble contacts with bound substrate. Corroborated by biochemical experiments and functional studies in yeast, we show that aliphatic residues flanking Tyr257 and Tyr662 are equally important for substrate interaction, and abolish Hsp104 function when mutated to glycine. Unexpectedly, we find that loop-2 is sensitive to aspartate substitutions that impair Hsp104 function and abolish protein disaggregation when loop-2 is replaced by four aspartate residues. Our observations suggest that Hsp104 pore loops have non-overlapping functions in protein disaggregation and together coordinate substrate binding, unfolding, and translocation through the Hsp104 hexamer.},
doi = {10.1042/BSR20171399},
journal = {Bioscience Reports},
number = 6,
volume = 37,
place = {United States},
year = {Fri Dec 22 00:00:00 EST 2017},
month = {Fri Dec 22 00:00:00 EST 2017}
}
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