Covalently linked HslU hexamers support a probabilistic mechanism that links ATP hydrolysis to protein unfolding and translocation

Baytshtok, Vladimir; Chen, Jiejin; Glynn, Steven E.; Nager, Andrew R.; Grant, Robert A.; Baker, Tania A.; Sauer, Robert T.

doi:10.1074/jbc.M116.768978

Covalently linked HslU hexamers support a probabilistic mechanism that links ATP hydrolysis to protein unfolding and translocation

Journal Article · Tue Feb 21 00:00:00 EST 2017 · Journal of Biological Chemistry

DOI:https://doi.org/10.1074/jbc.M116.768978· OSTI ID:1430301

Baytshtok, Vladimir ^[1]; Chen, Jiejin ^[2]; Glynn, Steven E. ^[3]; Nager, Andrew R. ^[4]; Grant, Robert A. ^[1]; Baker, Tania A. ^[1]; Sauer, Robert T. ^[1]

Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States)
Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Takeda Pharmaceuticals, Cambridge, MA (United States)
Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Stony Brook Univ., NY (United States)
Massachusetts Inst. of Technology (MIT), Cambridge, MA (United States); Stanford Univ., CA (United States)

The HslUV proteolytic machine consists of HslV, a double-ring self-compartmentalized peptidase, and one or two AAA+ HslU ring hexamers that hydrolyze ATP to power the unfolding of protein substrates and their translocation into the proteolytic chamber of HslV. Furthermore, we use genetic tethering and disulfide bonding strategies to construct HslU pseudohexamers containing mixtures of ATPase active and inactive subunits at defined positions in the hexameric ring. Genetic tethering impairs HslV binding and degradation, even for pseudohexamers with six active subunits, but disulfide-linked pseudohexamers do not have these defects, indicating that the peptide tether interferes with HslV interactions. Importantly, pseudohexamers containing different patterns of hydrolytically active and inactive subunits retain the ability to unfold protein substrates and/or collaborate with HslV in their degradation, supporting a model in which ATP hydrolysis and linked mechanical function in the HslU ring operate by a probabilistic mechanism.

View Accepted Manuscript (DOE)

Research Organization:: Argonne National Laboratory (ANL), Argonne, IL (United States)

Sponsoring Organization:: National Inst. of Health

OSTI ID:: 1430301

Journal Information:: Journal of Biological Chemistry, Journal Name: Journal of Biological Chemistry Journal Issue: 14 Vol. 292; ISSN 0021-9258

Publisher:: American Society for Biochemistry and Molecular BiologyCopyright Statement

Country of Publication:: United States

Language:: ENGLISH

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59 BASIC BIOLOGICAL SCIENCES
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protein unfolding

Covalently linked HslU hexamers support a probabilistic mechanism that links ATP hydrolysis to protein unfolding and translocation

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