title: A Mechanism of Subunit Recruitment in Human Small Heat Shock Protein Oligomers.
abstract: Small heat shock proteins (sHSPs) make up a class of molecular chaperones broadly observed across organisms. Many sHSPs form large oligomers that undergo dynamic subunit exchange that is thought to play a role in chaperone function. Though remarkably heterogeneous, sHSP oligomers share three types of intermolecular interactions that involve all three defined regions of a sHSP: the N-terminal region (NTR), the conserved α-crystallin domain (ACD), and a C-terminal region (CTR). Here we define the structural interactions involved in incorporation of a subunit into a sHSP oligomer. We demonstrate that a minimal ACD dimer of the human sHSP, HSPB5, interacts with an HSPB5 oligomer through two types of interactions: (1) interactions with CTRs in the oligomer and (2) via exchange into and out of the dimer interface composed of two ACDs. Unexpectedly, although dimers are thought to be the fundamental building block for sHSP oligomers, our results clearly indicate that subunit exchange into and out of oligomers occurs via monomers. Using structure-based mutants, we show that incorporation of a subunit into an oligomer is predicated on recruitment of the subunit via its interaction with CTRs on an oligomer. Both the rate and extent of subunit incorporation depend on the accessibility of CTRs within an HSPB5 oligomer. We show that this mechanism also applies to formation of heterooligomeric sHSP species composed of HSPB5 and HSPB6 and is likely general among sHSPs. Finally, our observations highlight the importance of NTRs in the thermodynamic stability of sHSP oligomers.
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195-7350, United States.
authors.forename: Scott P
authors.initials: SP
authors.lastname: Delbecq
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195-7350, United States.
authors.forename: Joel C
authors.initials: JC
authors.lastname: Rosenbaum
authors.affiliation: Department of Biochemistry, University of Washington, Seattle, Washington 98195-7350, United States.
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Klevit
mesh_terms: D050888:Heat-Shock Proteins, Small / Q000737:chemistry* / Q000378:metabolism; D006801:Humans; D008958:Models, Molecular; D011487:Protein Conformation; D054730:Protein Interaction Domains and Motifs; D055503:Protein Multimerization; D021122:Protein Subunits / Q000737:chemistry / Q000378:metabolism; D038201:alpha-Crystallins / Q000737:chemistry* / Q000378:metabolism
chemical_list: D050888:Heat-Shock Proteins, Small; D021122:Protein Subunits; D038201:alpha-Crystallins