Quaternary dynamics and plasticity underlie small heat shock protein chaperone function.
Small Heat Shock Proteins (sHSPs) are a diverse family of molecular chaperones that prevent protein aggregation by binding clients destabilized during cellular stress. Here we probe the architecture and dynamics of complexes formed between an oligomeric sHSP and client by employing unique mass spectrometry strategies. We observe over 300 different stoichiometries of interaction, demonstrating that an ensemble of structures underlies the protection these chaperones confer to unfolding clients. This astonishing heterogeneity not only makes the system quite distinct in behavior to ATP-dependent chaperones, but also renders it intractable by conventional structural biology approaches. We find that thermally regulated quaternary dynamics of the sHSP establish and maintain the plasticity of the system. This extends the paradigm that intrinsic dynamics are crucial to protein function to include equilibrium fluctuations in quaternary structure, and suggests they are integral to the sHSPs' role in the cellular protein homeostasis network.
Stengel
Florian
F
Department of Chemistry, University of Cambridge, Lensfield Road, Cambridge CB2 1EW, United Kingdom.
Baldwin
Andrew J
AJ
Painter
Alexander J
AJ
Jaya
Nomalie
N
Basha
Eman
E
Kay
Lewis E
LE
Vierling
Elizabeth
E
Robinson
Carol V
CV
Benesch
Justin L P
JL
D055592:Biophysical Phenomena; D006360:Heat-Shock Proteins / Q000737:chemistry / Q000378:metabolism; D050888:Heat-Shock Proteins, Small / Q000737:chemistry* / Q000378:metabolism*; D049409:Luciferases, Firefly / Q000737:chemistry / Q000378:metabolism; D008958:Models, Molecular; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism; D046912:Multiprotein Complexes; D018532:Pisum sativum / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry / Q000378:metabolism; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry; D013816:Thermodynamics
C489905:HSP18 protein, plant; D006360:Heat-Shock Proteins; D050888:Heat-Shock Proteins, Small; D018832:Molecular Chaperones; D046912:Multiprotein Complexes; D010940:Plant Proteins; D011994:Recombinant Proteins; C485197:luciferase, Photinus; D049409:Luciferases, Firefly